|
1
|
Wang X, Fang L, Xiao L, Zhong G, Han M, Wang B, Ren J, Zang Y. Research on the effect of LAMP1 in the development and progression of ccRCC and its potential mechanism with LC3C-mediated autophagy. Front Immunol 2024; 15:1494005. [PMID: 39669571 PMCID: PMC11634794 DOI: 10.3389/fimmu.2024.1494005] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2024] [Accepted: 11/11/2024] [Indexed: 12/14/2024] Open
Abstract
Background Lysomembrane-associated protein 1 (LAMP1), known to exhibit differential expression in various tumor types and play a crucial role in the development of tumors. Clear cell Renal Cell Carcinoma (ccRCC) is still the most common pathological type of renal carcinoma with poor prognosis. However, the expression of LAMP1 and its underlying molecular mechanism with ccRCC remain elusive. Methods Firstly, the expression of LAMP1 in ccRCC and its clinical significance were analyzed using various databases. Next, Weston Blot was performed to detect the expression of LAMP1 protein in cancer tissues and adjacent tissues from 60 pairs of clinical ccRCC patients. The correlation between LAMP1 expression and different clinical indicators as well as the relationship with patient prognosis was analyzed. Furthermore, molecular cell biology experiments were conducted to validate the effects of LAMP1 gene expression on cell proliferation, invasion and migration. Additionally, we investigated the impact of VHL, a key gene in renal cancer, and LC3C, an autophagy-related gene, on LAMP1 expression through molecular biology experiments to elucidate the potential underlying mechanism. Results Bioinformatics analysis revealed significant underexpression of LAMP1 in ccRCC (P<0.001), which correlated with poorer prognosis. In multivariate survival analysis, LAMP1 emerged as an independent prognostic marker for overall survival(OS)(P<0.05). Analysis of cancer and paracancer tissue samples from ccRCC patients demonstrated significantly lower levels of LAMP1 in tumors compared to paracancerous tissues (P<0.001), confirming its prognostic impact. Cell functionality experiment revealed that elevated LAMP1 inhibited cell proliferation, migration, and invasion. LAMP1 expression remained unchanged during autophagy modulation but decreased with LC3C knockdown and vice versa. Notably, VHL(+) cells expressed less LAMP1 than VHL(-) cells. Conclusions These findings indicate that low expression levels of LAMP1 is associated with poor prognosis in ccRCC. Therefore, LAMP1 emerges as a novel biomarker associated with the diagnosis and prognosis of renal cancer. Furthermore, we have also described the potential mechanism of action of LAMP1 in renal cancer. LAMP1 is a promising target for the treatment of ccRCC.
Collapse
Affiliation(s)
- Xiongbao Wang
- Department of Urology, Qilu Hospital of Shandong University, Jinan, Shandong, China
- Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, Shandong, China
- Laboratory of Basic Medical Sciences, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Liang Fang
- Department of Urology, Qilu Hospital of Shandong University, Jinan, Shandong, China
- Laboratory of Basic Medical Sciences, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Lixiang Xiao
- Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China
| | - Guangxin Zhong
- Department of Urology, School of Clinical Medicine, Beijing Tsinghua Changgung Hospital, Tsinghua University, Beijing, China
| | - Minghao Han
- Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China
| | - Bingshen Wang
- Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China
| | - Juchao Ren
- Department of Urology, Qilu Hospital of Shandong University, Jinan, Shandong, China
| | - Yuanwei Zang
- Department of Urology, Qilu Hospital of Shandong University, Jinan, Shandong, China
- Laboratory of Basic Medical Sciences, Qilu Hospital of Shandong University, Jinan, Shandong, China
| |
Collapse
|
|
2
|
Berg AL, Showalter MR, Kosaisawe N, Hu M, Stephens NC, Sa M, Heil H, Castro N, Chen JJ, VanderVorst K, Wheeler MR, Rabow Z, Cajka T, Albeck J, Fiehn O, Carraway KL. Cellular transformation promotes the incorporation of docosahexaenoic acid into the endolysosome-specific lipid bis(monoacylglycerol)phosphate in breast cancer. Cancer Lett 2023; 557:216090. [PMID: 36773796 PMCID: PMC10589064 DOI: 10.1016/j.canlet.2023.216090] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/23/2022] [Revised: 01/26/2023] [Accepted: 02/04/2023] [Indexed: 02/12/2023]
Abstract
Bis(monoacylglycero)phosphates (BMPs), a class of lipids highly enriched within endolysosomal organelles, are key components of the lysosomal intraluminal vesicles responsible for activating sphingolipid catabolic enzymes. While BMPs are understudied relative to other phospholipids, recent reports associate BMP dysregulation with a variety of pathological states including neurodegenerative diseases and lysosomal storage disorders. Since the dramatic lysosomal remodeling characteristic of cellular transformation could impact BMP abundance and function, we employed untargeted lipidomics approaches to identify and quantify BMP species in several in vitro and in vivo models of breast cancer and comparative non-transformed cells and tissues. We observed lower BMP levels within transformed cells relative to normal cells, and consistent enrichment of docosahexaenoic acid (22:6) fatty acyl chain-containing BMP species in both human- and mouse-derived mammary tumorigenesis models. Our functional analysis points to a working model whereby 22:6 BMPs serve as reactive oxygen species scavengers in tumor cells, protecting lysosomes from oxidant-induced lysosomal membrane permeabilization. Our findings suggest that breast tumor cells might divert polyunsaturated fatty acids into BMP lipids as part of an adaptive response to protect their lysosomes from elevated reactive oxygen species levels, and raise the possibility that BMP-mediated lysosomal protection is a tumor-specific vulnerability that may be exploited therapeutically.
Collapse
Affiliation(s)
- Anastasia L Berg
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA
| | - Megan R Showalter
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA
| | - Nont Kosaisawe
- Department of Molecular and Cellular Biology, University of California Davis, Davis, CA, USA
| | - Michelle Hu
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA
| | - Nathanial C Stephens
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA
| | - Michael Sa
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA
| | - Hailey Heil
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA
| | - Noemi Castro
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA
| | - Jenny J Chen
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA
| | - Kacey VanderVorst
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA
| | - Madelyn R Wheeler
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA
| | - Zachary Rabow
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA
| | - Tomas Cajka
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA; Institute of Physiology of the Czech Academy of Sciences, Prague, 14200, Czech Republic
| | - John Albeck
- Department of Molecular and Cellular Biology, University of California Davis, Davis, CA, USA
| | - Oliver Fiehn
- West Coast Metabolomics Center, UC Davis Genome Center, University of California Davis, Davis, CA, USA
| | - Kermit L Carraway
- Department of Biochemistry and Molecular Medicine and UC Davis Comprehensive Cancer Center, University of California Davis School of Medicine, Sacramento, CA, USA.
| |
Collapse
|
|
3
|
Berg AL, Rowson-Hodel A, Wheeler MR, Hu M, Free SR, Carraway KL. Engaging the Lysosome and Lysosome-Dependent Cell Death in Cancer. Breast Cancer 2022. [DOI: 10.36255/exon-publications-breast-cancer-lysosome] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
|
|
4
|
Capone E, Iacobelli S, Sala G. Role of galectin 3 binding protein in cancer progression: a potential novel therapeutic target. J Transl Med 2021; 19:405. [PMID: 34565385 PMCID: PMC8474792 DOI: 10.1186/s12967-021-03085-w] [Citation(s) in RCA: 78] [Impact Index Per Article: 19.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/14/2021] [Accepted: 09/16/2021] [Indexed: 12/19/2022] Open
Abstract
The lectin galactoside-binding soluble 3 binding protein (LGALS3BP) is a secreted, hyperglycosylated protein expressed by the majority of human cells. It was first identified as cancer and metastasis associated protein, while its role in innate immune response upon viral infection remains still to be clarified. Since its discovery dated in early 90 s, a large body of literature has been accumulating highlighting both a prognostic and functional role for LGALS3BP in cancer. Moreover, data from our group and other have strongly suggested that this protein is enriched in cancer-associated extracellular vesicles and may be considered a promising candidate for a targeted therapy in LGALS3BP positive cancers. Here, we extensively reviewed the literature relative to LGALS3BP role in cancer and its potential value as a therapeutic target.
Collapse
Affiliation(s)
- Emily Capone
- Department of Innovative Technologies in Medicine and Dentistry, University of Chieti-Pescara, 66100, Chieti, Italy.,Center for Advanced Studies and Technology (CAST), Via Polacchi 11, 66100, Chieti, Italy
| | | | - Gianluca Sala
- Department of Innovative Technologies in Medicine and Dentistry, University of Chieti-Pescara, 66100, Chieti, Italy. .,Center for Advanced Studies and Technology (CAST), Via Polacchi 11, 66100, Chieti, Italy.
| |
Collapse
|
|
5
|
Luo M, Zhang Q, Hu Y, Sun C, Sheng Y, Deng C. LGALS3BP: A Potential Plasma Biomarker Associated with Diagnosis and Prognosis in Patients with Sepsis. Infect Drug Resist 2021; 14:2863-2871. [PMID: 34335032 PMCID: PMC8318715 DOI: 10.2147/idr.s316402] [Citation(s) in RCA: 13] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/22/2021] [Accepted: 06/24/2021] [Indexed: 12/13/2022] Open
Abstract
Purpose This study aimed to screen differentially expressed proteins (DEPs) in plasma of patients with sepsis through data-independent acquisition (DIA) and enzyme-linked immunosorbent assays (ELISAs), and provide convenient and accurate serum markers for determining the condition of septic patients. Methods A total of 53 septic patients and 16 normal controls who were admitted to the Affiliated Hospital of Southwest Medical University between January 2019 and December 2020 were enrolled in this study; 6 specimens from the normal group and 15 from the sepsis group were randomly selected for DIA-based quantitative proteomic analysis. The acquired data were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and a protein-protein interaction (PPI) network was constructed to screen potential markers. The selected proteins were further verified through ELISAs. The differences between control and sepsis groups and between survivors and non-survivors were analysed. Receiver operating characteristic (ROC) curves were drawn to explore their diagnostic value and prognostic efficacy. Results A total of 149 DEPs were identified by bioinformatics methods. The analyses showed that these proteins are mainly involved in biological processes such as cell movement, stress response, cell proliferation, and immune response. Functional pathway analysis showed that they are mainly involved in leukocyte transendothelial migration, protein synthesis and processing, and various bacterial infections. LGALS3BP was selected as a potential plasma biomarker and further verified through an ELISA. Its level in septic patients was significantly higher than that in normal controls, and its level in non-survivors was also higher than that in survivors. The ROC curves suggested its great diagnostic efficacy and prognostic ability in sepsis. Conclusion LGALS3BP levels were significantly different between the normal and sepsis groups; it has good diagnostic value in sepsis, and is related to patient prognosis; thus, it might be a biomarker for sepsis.
Collapse
Affiliation(s)
- Meiyan Luo
- Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Department of Tuberculosis, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Infection and Immunity Laboratory,The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China
| | - Qian Zhang
- Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Department of Tuberculosis, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Infection and Immunity Laboratory,The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China
| | - Yingchun Hu
- Department of Emergency, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China
| | - Changfeng Sun
- Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Department of Tuberculosis, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Infection and Immunity Laboratory,The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China
| | - Yunjian Sheng
- Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Department of Tuberculosis, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Infection and Immunity Laboratory,The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China
| | - Cunliang Deng
- Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Department of Tuberculosis, The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China.,Infection and Immunity Laboratory,The Affiliated Hospital of Southwest Medical University, Louzhou, 646000, People's Republic of China
| |
Collapse
|
|
6
|
Cho SH, Shim HJ, Park MR, Choi JN, Akanda MR, Hwang JE, Bae WK, Lee KH, Sun EG, Chung IJ. Lgals3bp suppresses colon inflammation and tumorigenesis through the downregulation of TAK1-NF-κB signaling. Cell Death Discov 2021; 7:65. [PMID: 33824294 PMCID: PMC8024364 DOI: 10.1038/s41420-021-00447-7] [Citation(s) in RCA: 19] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/02/2020] [Revised: 02/18/2021] [Accepted: 03/08/2021] [Indexed: 12/21/2022] Open
Abstract
Galectin 3-binding protein (LGALS3BP, also known as 90K) is a multifunctional glycoprotein involved in immunity and cancer. However, its precise role in colon inflammation and tumorigenesis remains unclear. Here, we showed that Lgals3bp-/- mice were highly susceptible to colitis and colon tumorigenesis, accompanied by the induction of inflammatory responses. In acute colitis, NF-κB was highly activated in the colon of Lgals3bp-/- mice, leading to the excessive production of pro-inflammatory cytokines, such as IL-6, TNFα, and IL-1β. Mechanistically, Lgals3bp suppressed NF-κB through the downregulation of TAK1 in colon epithelial cells. There was no significant difference in the pro-inflammatory cytokine levels between wild-type and Lgals3bp-/- mice in a chronic inflammatory state, during colon tumorigenesis. Instead, Lgals3bp-/- mice showed elevated levels of GM-CSF, compared to those in WT mice. We also found that GM-CSF promoted the accumulation of myeloid-derived suppressor cells and ultimately increased colon tumorigenesis in Lgals3bp-/- mice. Taken together, Lgals3bp plays a critical role in the suppression of colitis and colon tumorigenesis through the downregulation of the TAK1-NF-κB-cytokine axis. These findings suggest that LGALS3BP is a novel immunotherapeutic target for colon inflammation and tumorigenesis.
Collapse
Affiliation(s)
- Sang-Hee Cho
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
- Immunotherapy Innovation Center, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
| | - Hyun-Jeong Shim
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
| | - Mi-Ra Park
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
| | - Ji-Na Choi
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
| | - Md Rashedunnabi Akanda
- Combinatorial Tumor Immunotherapy MRC Center, Chonnam National University Medical School, Hwasun, Republic of Korea
- Department of Pathology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
- Department of Pharmacology and Toxicology, Sylhet Agricultural University, Sylhet, Bangladesh
| | - Jun-Eul Hwang
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
| | - Woo-Kyun Bae
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
- Combinatorial Tumor Immunotherapy MRC Center, Chonnam National University Medical School, Hwasun, Republic of Korea
| | - Kyung-Hwa Lee
- Immunotherapy Innovation Center, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
- Combinatorial Tumor Immunotherapy MRC Center, Chonnam National University Medical School, Hwasun, Republic of Korea
- Department of Pathology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea
| | - Eun-Gene Sun
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea.
| | - Ik-Joo Chung
- Department of Hematology and Oncology, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea.
- Immunotherapy Innovation Center, Chonnam National University Medical School and Hwasun Hospital, Hwasun, Republic of Korea.
| |
Collapse
|
|
7
|
Lin HZ, Zhang T, Chen MY, Shen JL. Novel biomarkers for the diagnosis and prognosis of gallbladder cancer. J Dig Dis 2021; 22:62-71. [PMID: 33369216 DOI: 10.1111/1751-2980.12966] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/14/2020] [Revised: 11/10/2020] [Accepted: 12/22/2020] [Indexed: 01/17/2023]
Abstract
Gallbladder cancer (GBC) is the most common form of biliary tract malignancy with a dismal prognosis. A poor outcome in patients with GBC is related to the aggressive nature of the tumor, delayed diagnosis, and a lack of reliable biomarkers and effective treatment. Therefore, early diagnosis and accurate disease assessment are crucial to prolonging the patient survival. Identification of novel prognostic and diagnostic biomarkers may help improve the early diagnostic rate and develop specific targeted treatments for patients with GBC. We herein review the novel biomarkers that may be associated with the diagnosis and prognosis in GBC and their potential clinical significance in the management of GBC.
Collapse
Affiliation(s)
- Hong Ze Lin
- Nanshan School, Guangzhou Medical University, Guangzhou, Guangdong Province, China
| | - Tao Zhang
- Nanshan School, Guangzhou Medical University, Guangzhou, Guangdong Province, China
| | - Ming Yu Chen
- Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
| | - Ji Liang Shen
- Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, China
| |
Collapse
|
|
8
|
Ferdoushi A, Li X, Griffin N, Faulkner S, Jamaluddin MFB, Gao F, Jiang CC, van Helden DF, Tanwar PS, Jobling P, Hondermarck H. Schwann Cell Stimulation of Pancreatic Cancer Cells: A Proteomic Analysis. Front Oncol 2020; 10:1601. [PMID: 32984024 PMCID: PMC7477957 DOI: 10.3389/fonc.2020.01601] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/19/2020] [Accepted: 07/23/2020] [Indexed: 12/12/2022] Open
Abstract
Schwann cells (SCs), the glial component of peripheral nerves, have been identified as promoters of pancreatic cancer (PC) progression, but the molecular mechanisms are unclear. In the present study, we aimed to identify proteins released by SCs that could stimulate PC growth and invasion. Proteomic analysis of human primary SC secretome was performed using liquid chromatography–tandem mass spectrometry, and a total of 13,796 unique peptides corresponding to 1,470 individual proteins were identified. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment were conducted using the Database for Annotation, Visualization, and Integrated Discovery. Metabolic and cell–cell adhesion pathways showed the highest levels of enrichment, a finding in line with the supportive role of SCs in peripheral nerves. We identified seven SC-secreted proteins that were validated by western blot. The involvement of these SC-secreted proteins was further demonstrated by using blocking antibodies. PC cell proliferation and invasion induced by SC-conditioned media were decreased using blocking antibodies against the matrix metalloproteinase-2, cathepsin D, plasminogen activator inhibitor-1, and galectin-1. Blocking antibodies against the proteoglycan biglycan, galectin-3 binding protein, and tissue inhibitor of metalloproteinases-2 decreased only the proliferation but not the invasion of PC cells. Together, this study delineates the secretome of human SCs and identifies proteins that can stimulate PC cell growth and invasion and therefore constitute potential therapeutic targets.
Collapse
Affiliation(s)
- Aysha Ferdoushi
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia.,Department of Biotechnology and Genetic Engineering, Mawlana Bhashani Science and Technology University, Tangail, Bangladesh
| | - Xiang Li
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Nathan Griffin
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Sam Faulkner
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - M Fairuz B Jamaluddin
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Fangfang Gao
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Chen Chen Jiang
- Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia.,School of Medicine and Public Health, The University of Newcastle, Callaghan, NSW, Australia
| | - Dirk F van Helden
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Pradeep S Tanwar
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Phillip Jobling
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| | - Hubert Hondermarck
- School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.,Hunter Medical Research Institute, University of Newcastle, New Lambton, NSW, Australia
| |
Collapse
|
|
9
|
Samonig L, Loipetzberger A, Blöchl C, Rurik M, Kohlbacher O, Aberger F, Huber CG. Proteins and Molecular Pathways Relevant for the Malignant Properties of Tumor-Initiating Pancreatic Cancer Cells. Cells 2020; 9:E1397. [PMID: 32503348 PMCID: PMC7349116 DOI: 10.3390/cells9061397] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/30/2020] [Revised: 05/26/2020] [Accepted: 05/30/2020] [Indexed: 12/29/2022] Open
Abstract
Cancer stem cells (CSCs), a small subset of the tumor bulk with highly malignant properties, are deemed responsible for tumor initiation, growth, metastasis, and relapse. In order to reveal molecular markers and determinants of their tumor-initiating properties, we enriched rare stem-like pancreatic tumor-initiating cells (TICs) by harnessing their clonogenic growth capacity in three-dimensional multicellular spheroid cultures. We compared pancreatic TICs isolated from three-dimensional tumor spheroid cultures with nontumor-initiating cells (non-TICs) enriched in planar cultures. Employing differential proteomics (PTX), we identified more than 400 proteins with significantly different expression in pancreatic TICs and the non-TIC population. By combining the unbiased PTX with mRNA expression analysis and literature-based predictions of pro-malignant functions, we nominated the two calcium-binding proteins S100A8 (MRP8) and S100A9 (MRP14) as well as galactin-3-binding protein LGALS3BP (MAC-2-BP) as putative determinants of pancreatic TICs. In silico pathway analysis followed by candidate-based RNA interference mediated loss-of-function analysis revealed a critical role of S100A8, S100A9, and LGALS3BP as molecular determinants of TIC proliferation, migration, and in vivo tumor growth. Our study highlights the power of combining unbiased proteomics with focused gene expression and functional analyses for the identification of novel key regulators of TICs, an approach that warrants further application to identify proteins and pathways amenable to drug targeting.
Collapse
Affiliation(s)
- Lisa Samonig
- Department of Biosciences, Bioanalytical Research Labs, University of Salzburg, A-5020 Salzburg, Austria; (L.S.); (C.B.)
| | - Andrea Loipetzberger
- Department of Biosciences, Cancer Cluster Salzburg, Molecular Cancer and Stem Cell Research, University of Salzburg, A-5020 Salzburg, Austria;
| | - Constantin Blöchl
- Department of Biosciences, Bioanalytical Research Labs, University of Salzburg, A-5020 Salzburg, Austria; (L.S.); (C.B.)
| | - Marc Rurik
- Institute for Bioinformatics and Medical Informatics, University of Tübingen, Sand 14, 72076 Tübingen, Germany; (M.R.); (O.K.)
| | - Oliver Kohlbacher
- Institute for Bioinformatics and Medical Informatics, University of Tübingen, Sand 14, 72076 Tübingen, Germany; (M.R.); (O.K.)
- Biomolecular Interactions, Max Planck Institute for Developmental Biology, Max-Planck-Ring 5, 72076 Tübingen, Germany
- Institute for Translational Bioinformatics, University Hospital Tübingen, Hoppe-Seyler-Str. 9, 72076 Tübingen, Germany
- Quantitative Biology Center, University of Tübingen, Auf der Morgenstelle 10, 72076 Tübingen, Germany
| | - Fritz Aberger
- Department of Biosciences, Cancer Cluster Salzburg, Molecular Cancer and Stem Cell Research, University of Salzburg, A-5020 Salzburg, Austria;
- Department of Biosciences, Cancer Cluster Salzburg, University of Salzburg, A-5020 Salzburg, Austria
| | - Christian G. Huber
- Department of Biosciences, Bioanalytical Research Labs, University of Salzburg, A-5020 Salzburg, Austria; (L.S.); (C.B.)
- Department of Biosciences, Cancer Cluster Salzburg, University of Salzburg, A-5020 Salzburg, Austria
| |
Collapse
|
|
10
|
Abeywickrama CS, Wijesinghe KJ, Stahelin RV, Pang Y. Lysosome imaging in cancer cells by pyrene-benzothiazolium dyes: An alternative imaging approach for LAMP-1 expression based visualization methods to avoid background interference. Bioorg Chem 2019; 91:103144. [PMID: 31377388 PMCID: PMC7065667 DOI: 10.1016/j.bioorg.2019.103144] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/30/2019] [Revised: 07/03/2019] [Accepted: 07/19/2019] [Indexed: 12/31/2022]
Abstract
A series of pyrene-benzothiazolium dyes (1a-1d) were experimentally investigated to study their internalization mechanism into cellular lysosomes as well as their potential imaging applications for live cell imaging. The lysosome selectivity of the probes was further compared by using fluorescently tagged lysosome associated membrane protein-1 (LAMP-1) expression-dependent visualization in both normal (COS-7, HEK293) and cancer (A549, Huh 7.5) cell lines. These probes were successfully employed as reliable lysosome markers in tumor cell models, thus providing an attractive alternative to LAMP-1 expression-dependent visualization methods. One advantage of these probes is the elimination of significant background fluorescence arising from fluorescently tagged protein expression on the cell surface when cells were transfected with LAMP-1 expression plasmids. Probes exhibited remarkable ability to stain cellular lysosomes for long-term experiments (up to 24 h) and the highly lipophilic nature of the probe design allowed their accumulation in hydrophobic regions of the cellular lysosomes. Experimental evidences indicated that the probes are likely to be internalized into lysosomes via endocytosis and accumulated in the hydrophobic regions of the lysosomes rather than in the acidic lysosomal lumen. These probes also demonstrated significant stability and lysosome staining for fixed cell imaging applications as well. Lastly, the benzothiazolium moiety of the probes was identified as the key component for lysosome selectivity.
Collapse
Affiliation(s)
| | - Kaveesha J Wijesinghe
- Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA
| | - Robert V Stahelin
- Department of Medicinal Chemistry and Molecular Pharmacology and the Purdue Center for Cancer Research, Purdue University, West Lafayette, IN 47907, USA.
| | - Yi Pang
- Department of Chemistry, University of Akron, Akron, OH 44325, USA; Maurice Morton Institute of Polymer Science, University of Akron, Akron, OH 44325, USA.
| |
Collapse
|
|
11
|
Duangkumpha K, Stoll T, Phetcharaburanin J, Yongvanit P, Thanan R, Techasen A, Namwat N, Khuntikeo N, Chamadol N, Roytrakul S, Mulvenna J, Mohamed A, Shah AK, Hill MM, Loilome W. Urine proteomics study reveals potential biomarkers for the differential diagnosis of cholangiocarcinoma and periductal fibrosis. PLoS One 2019; 14:e0221024. [PMID: 31425520 PMCID: PMC6699711 DOI: 10.1371/journal.pone.0221024] [Citation(s) in RCA: 20] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2019] [Accepted: 07/30/2019] [Indexed: 12/15/2022] Open
Abstract
Cholangiocarcinoma (CCA) is a primary malignant tumor of the epithelial lining of biliary track associated with endemic Opisthorchis viverrini (Ov) infection in northeastern Thailand. Ov-associated periductal fibrosis (PDF) is the precancerous lesion for CCA, and can be detected by ultrasonography (US) to facilitate early detection. However, US cannot be used to distinguish PDF from cancer. Therefore, the objective of this study was to discover and qualify potential urine biomarkers for CCA detection in at-risk population. Biomarker discovery was conducted on pooled urine samples, 42 patients per group, with PDF or normal bile duct confirmed by ultrasound. After depletion of high abundance proteins, 338 urinary proteins were identified from the 3 samples (normal-US, PDF-US, CCA). Based on fold change and literature review, 70 candidate proteins were selected for qualification by multiple reaction monitoring mass spectrometry (MRM-MS) in 90 individual urine samples, 30 per group. An orthogonal signal correction projection to latent structures discriminant analysis (O-PLS-DA) multivariate model constructed from the 70 candidate biomarkers significantly discriminated CCA from normal and PDF groups (P = 0.003). As an independent validation, the expression of 3 candidate proteins was confirmed by immunohistochemistry in CCA tissues: Lysosome associated membrane glycoprotein 1 (LAMP1), lysosome associated membrane glycoprotein 2 (LAMP2) and cadherin-related family member 2 (CDHR2). Further evaluation of these candidate biomarkers in a larger cohort is needed to support their applicability in a clinical setting for screening and monitoring early CCA and for CCA surveillance.
Collapse
Affiliation(s)
- Kassaporn Duangkumpha
- Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.,Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand
| | - Thomas Stoll
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Jutarop Phetcharaburanin
- Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.,Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand
| | - Puangrat Yongvanit
- Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand
| | - Raynoo Thanan
- Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand
| | - Anchalee Techasen
- Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand.,Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand
| | - Nisana Namwat
- Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.,Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand
| | - Narong Khuntikeo
- Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand.,Department of Surgery, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand
| | - Nittaya Chamadol
- Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand.,Department of Radiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand
| | - Sittiruk Roytrakul
- Proteomics Research Laboratory, Genome Institute, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani, Thailand
| | - Jason Mulvenna
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Ahmed Mohamed
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Alok K Shah
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Michelle M Hill
- QIMR Berghofer Medical Research Institute, Brisbane, Australia
| | - Watcharin Loilome
- Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.,Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand
| |
Collapse
|
|
12
|
Hong CS, Park MR, Sun EG, Choi W, Hwang JE, Bae WK, Rhee JH, Cho SH, Chung IJ. Gal-3BP Negatively Regulates NF-κB Signaling by Inhibiting the Activation of TAK1. Front Immunol 2019; 10:1760. [PMID: 31402917 PMCID: PMC6677151 DOI: 10.3389/fimmu.2019.01760] [Citation(s) in RCA: 21] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/17/2019] [Accepted: 07/11/2019] [Indexed: 01/02/2023] Open
Abstract
Galectin-3-binding protein (Gal-3BP) is a member of the family of scavenger receptor cysteine-rich (SRCR) domain-containing proteins, which are associated with the immune system. However, the functional roles and signaling mechanisms of Gal-3BP in host defense and the immune response remain largely unknown. Here, we identified cellular Gal-3BP as a negative regulator of NF-κB activation and proinflammatory cytokine production in lipopolysaccharide (LPS)-stimulated murine embryonic fibroblasts (MEFs). Furthermore, cellular Gal-3BP interacted with transforming growth factor β-activated kinase 1 (TAK1), a crucial mediator of NF-κB activation in response to cellular stress. Gal-3BP inhibited the phosphorylation of TAK1, leading to suppression of its kinase activity and reduced protein stability. In vivo we found that Lgals3BP deficiency in mice enhanced LPS-induced proinflammatory cytokine release and rendered mice more sensitive to LPS-induced endotoxin shock. Overall, these results suggest that Gal-3BP is a novel suppressor of TAK1-dependent NF-κB activation that may have potential in the prevention and treatment of inflammatory diseases.
Collapse
Affiliation(s)
- Chang-Soo Hong
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea
| | - Mi-Ra Park
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea
| | - Eun-Gene Sun
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea
| | - Wonyoung Choi
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea
| | - Jun-Eul Hwang
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea
| | - Woo-Kyun Bae
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea.,Combinatorial Tumor Immunotherapy MRC, Clinical Vaccine R&D Center and Department of Microbiology, Chonnam National University Medical School, Hwasun, South Korea
| | - Joon Haeng Rhee
- Combinatorial Tumor Immunotherapy MRC, Clinical Vaccine R&D Center and Department of Microbiology, Chonnam National University Medical School, Hwasun, South Korea
| | - Sang-Hee Cho
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea
| | - Ik-Joo Chung
- Department of Internal Medicine, Chonnam National University Medical School, Hwasun, South Korea.,Combinatorial Tumor Immunotherapy MRC, Clinical Vaccine R&D Center and Department of Microbiology, Chonnam National University Medical School, Hwasun, South Korea
| |
Collapse
|
|
13
|
Zhang X, Ding H, Lu Z, Ding L, Song Y, Jing Y, Hu Q, Dong Y, Ni Y. Increased LGALS3BP promotes proliferation and migration of oral squamous cell carcinoma via PI3K/AKT pathway. Cell Signal 2019; 63:109359. [PMID: 31302247 DOI: 10.1016/j.cellsig.2019.109359] [Citation(s) in RCA: 20] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/12/2019] [Revised: 07/04/2019] [Accepted: 07/10/2019] [Indexed: 12/18/2022]
Abstract
Previous studies showed that lectin galactoside-binding soluble 3 binding protein (LGALS3BP) is an important participant in tumor progression. However, its prognostic value and functional mechanism in oral squamous cell carcinoma (OSCC) are still unclear. In this study, we analyzed LGALS3BP expression in OSCC tissues via Oncomine databases and immunohistochemical staining. LGALS3BP was significantly up-regulated in OSCC tumor tissues. IHC analysis showed that LGALS3BP was predominantly expressed in tumor cells and correlated with poor clinical characteristics. In addition, high LGALS3BP expression predicted poor clinical outcomes and multivariate analysis revealed that LGALS3BP expression was as an independent prognostic factor for OS, DFS and RFS (p < .0001, p = .002, p = .002). Mechanically, LGALS3BP regulated OSCC proliferation and migration via PI3K/AKT pathways, which was abrogated by PI3K inhibitor LY294002 in a dose-dependent manner. Our results suggested that LGALS3BP could be served as a novel independent prognostic factor as well as a potential therapeutic target for OSCC treatment.
Collapse
Affiliation(s)
- Xiaoxin Zhang
- Central Laboratory, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Haoyue Ding
- Department of Oral and Maxillofacial Surgery, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Zhanyi Lu
- Department of Oral and Maxillofacial Surgery, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Liang Ding
- Central Laboratory, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Yuxian Song
- Central Laboratory, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Yue Jing
- Central Laboratory, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Qingang Hu
- Department of Oral and Maxillofacial Surgery, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China
| | - Yingchun Dong
- Department of Anesthesiology, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China.
| | - Yanhong Ni
- Central Laboratory, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, China.
| |
Collapse
|
|
14
|
Giansanti F, Capone E, Ponziani S, Piccolo E, Gentile R, Lamolinara A, Di Campli A, Sallese M, Iacobelli V, Cimini A, De Laurenzi V, Lattanzio R, Piantelli M, Ippoliti R, Sala G, Iacobelli S. Secreted Gal-3BP is a novel promising target for non-internalizing Antibody-Drug Conjugates. J Control Release 2018; 294:176-184. [PMID: 30553852 DOI: 10.1016/j.jconrel.2018.12.018] [Citation(s) in RCA: 26] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/16/2018] [Revised: 12/11/2018] [Accepted: 12/12/2018] [Indexed: 12/16/2022]
Abstract
Galectin-3-binding protein (Gal-3BP) has been identified as a cancer and metastasis-associated, secreted protein that is expressed by the large majority of cancers. The present study describes a special type of non-internalizing antibody-drug-conjugates that specifically target Gal-3BP. Here, we show that the humanized 1959 antibody, which specifically recognizes secreted Gal-3BP, selectively localized around tumor but not normal cells. A site specific disulfide linkage with thiol-maytansinoids to unpaired cysteine residues of 1959, resulting in a drug-antibody ratio of 2, yielded an ADC product, which cured A375m melanoma bearing mice. ADC products based on the non-internalizing 1959 antibody may be useful for the treatment of several human malignancies, as the cognate antigen is abundantly expressed and secreted by several cancers, while being present at low levels in most normal adult tissues.
Collapse
Affiliation(s)
| | - Emily Capone
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, Chieti, Italy
| | - Sara Ponziani
- Department MESVA, University of L'Aquila, 67100 Coppito, Italy; MediaPharma s.r.l., Via della Colonnetta 50/A, 66100 Chieti, Italy
| | - Enza Piccolo
- MediaPharma s.r.l., Via della Colonnetta 50/A, 66100 Chieti, Italy
| | - Roberta Gentile
- MediaPharma s.r.l., Via della Colonnetta 50/A, 66100 Chieti, Italy
| | - Alessia Lamolinara
- Department of Medicine and Aging Cesi-Met, Via Polacchi 11, 66100 Chieti, Italy
| | - Antonella Di Campli
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, Chieti, Italy
| | - Michele Sallese
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, Chieti, Italy
| | - Valentina Iacobelli
- Department of Gynecology & Obstetrics, Sapienza University of Rome, 00100 Rome, Italy
| | | | - Vincenzo De Laurenzi
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, Chieti, Italy
| | - Rossano Lattanzio
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, Chieti, Italy
| | - Mauro Piantelli
- MediaPharma s.r.l., Via della Colonnetta 50/A, 66100 Chieti, Italy
| | | | - Gianluca Sala
- Department of Medical, Oral and Biotechnological Sciences, University of Chieti-Pescara, Chieti, Italy; MediaPharma s.r.l., Via della Colonnetta 50/A, 66100 Chieti, Italy.
| | | |
Collapse
|
|
15
|
Dang Q, Zhou H, Qian J, Yang L, Huang J, Zhang Y, Shi W. LAMP1 Overexpression Predicts for Poor Prognosis in Diffuse Large B-cell Lymphoma. CLINICAL LYMPHOMA, MYELOMA & LEUKEMIA 2018; 18:749-754. [PMID: 30082222 DOI: 10.1016/j.clml.2018.07.288] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/02/2018] [Revised: 06/28/2018] [Accepted: 07/09/2018] [Indexed: 12/22/2022]
Abstract
INTRODUCTION Lysosomal-associated membrane protein 1 (LAMP1) is a lysosomal and plasma membrane protein that contributes to tumor metastatic potential and differentiation. PATIENTS AND METHODS We performed immunohistochemical staining to investigate LAMP1 protein expression levels in 122 diffuse large B-cell lymphoma (DLBCL) tumor samples and 45 reactive hyperplasia tissues. Correlations between LAMP1 expression, various clinicopathologic features, and patient prognosis were evaluated by univariate and multivariate analyses. RESULTS LAMP1 expression was greater in the DLBCL tissues than in the reactive hyperplasia tissues. High LAMP1 expression was significantly associated with a high international prognostic index (score, 3-5; P = .023) and elevated lactate dehydrogenase level (P = .028). Moreover, high LAMP1 expression (P = .026), elevated serum lactate dehydrogenase level (P = .011), and high international prognostic index (P < .001) were independently associated with worse overall survival and progression-free survival. CONCLUSION These data provide the first evidence that LAMP1 expression is associated with a poor prognosis in patients with DLBCL.
Collapse
Affiliation(s)
- Qingxiu Dang
- Department of Hematology, Affiliated Hospital of Nantong University, Nantong, China
| | - Hong Zhou
- Department of Hematology, Affiliated Hospital of Nantong University, Nantong, China
| | - Juan Qian
- Department of Hematology, Affiliated Hospital of Nantong University, Nantong, China
| | - Li Yang
- Department of Hematology, Affiliated Hospital of Nantong University, Nantong, China
| | - Jianfei Huang
- Clinical Biobank, Department of Pathology, Affiliated Hospital of Nantong University, Nantong, China
| | - Yaping Zhang
- Department of Hematology, Affiliated Hospital of Nantong University, Nantong, China.
| | - Wenyu Shi
- Department of Hematology, Affiliated Hospital of Nantong University, Nantong, China.
| |
Collapse
|
|
16
|
Zuo C, Sheng X, Ma M, Xia M, Ouyang L. ISG15 in the tumorigenesis and treatment of cancer: An emerging role in malignancies of the digestive system. Oncotarget 2018; 7:74393-74409. [PMID: 27626310 PMCID: PMC5342061 DOI: 10.18632/oncotarget.11911] [Citation(s) in RCA: 38] [Impact Index Per Article: 5.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/10/2016] [Accepted: 09/01/2016] [Indexed: 02/07/2023] Open
Abstract
The interferon-stimulated gene 15 ubiquitin-like modifier (ISG15) encodes an IFN-inducible, ubiquitin-like protein. The ISG15 protein forms conjugates with numerous cellular proteins that are involved in a multitude of cellular functions, including interferon-induced immune responses and the regulation of cellular protein turnover. The expression of ISG15 and ISG15-mediated conjugation has been implicated in a wide range of human tumors and cancer cell lines, but the roles of ISG15 in tumorigenesis and responses to anticancer treatments remain largely unknown. In this review, we discuss the findings of recent studies with regard to the role of ISG15 pathways in cancers of the digestive system.
Collapse
Affiliation(s)
- Chaohui Zuo
- Department of Gastroduodenal and Pancreatic Surgery, Translation Medicine Research Center of Liver Cancer, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, Hunan, China.,Graduate School, University of South China, Hengyang, Hunan, China
| | - Xinyi Sheng
- Graduate School, University of South China, Hengyang, Hunan, China
| | - Min Ma
- Department of Gastroduodenal and Pancreatic Surgery, Translation Medicine Research Center of Liver Cancer, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, Hunan, China
| | - Man Xia
- Laboratory of Digestive Oncology, Hunan Province Cancer Institute, Changsha, Hunan, China
| | - Linda Ouyang
- Laboratory of Digestive Oncology, Hunan Province Cancer Institute, Changsha, Hunan, China
| |
Collapse
|
|
17
|
Yumioka T, Osaki M, Sasaki R, Yamaguchi N, Onuma K, Iwamoto H, Morizane S, Honda M, Takenaka A, Okada F. Lysosome-associated membrane protein 2 (LAMP-2) expression induced by miR-194-5p downregulation contributes to sunitinib resistance in human renal cell carcinoma cells. Oncol Lett 2017; 15:893-900. [PMID: 29399154 PMCID: PMC5772808 DOI: 10.3892/ol.2017.7423] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/08/2017] [Accepted: 11/07/2017] [Indexed: 12/31/2022] Open
Abstract
Sunitinib is a tyrosine kinase inhibitor that is used as the primary treatment in metastatic renal cell carcinoma (RCC). The main difficulty associated with its use is the development of drug resistance. In the present study, ACHN cells, a human renal cell carcinoma cell line, were used to establish sunitinib-resistant (SR) cells. Microarray analysis and reverse transcription-quantitative polymerase chain reaction revealed that miR-194-5p expression was significantly decreased in SR-ACHN cells when compared with that observed in ACHN cells (P<0.05). Transfection of miR-194-5p, though not with negative control miR, in SR-ACHN cells could significantly inhibit cell proliferation following sunitinib treatment (2.5–40 µM; P<0.05). Western blotting demonstrated that the expression of lysosome-associated membrane protein-2 (LAMP-2), which attenuates the anti-proliferative effect of sunitinib, was significantly higher in SR-ACHN than in ACHN cells (P<0.01). In addition, LAMP-2 expression was suppressed by miR-194-5p transfection in SR-ACHN cells. These data suggested that miR-194-5p downregulation may be associated with sunitinib resistance via the induction of LAMP-2 expression in human RCC.
Collapse
Affiliation(s)
- Tetsuya Yumioka
- Division of Pathological Biochemistry, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan.,Division of Urology, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Mitsuhiko Osaki
- Division of Pathological Biochemistry, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan.,Chromosome Engineering Research Center, Tottori University, Tottori 683-8503, Japan
| | - Ryo Sasaki
- Division of Pathological Biochemistry, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Noriya Yamaguchi
- Division of Pathological Biochemistry, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan.,Division of Urology, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Kunishige Onuma
- Division of Pathological Biochemistry, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Hideto Iwamoto
- Division of Urology, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Shuichi Morizane
- Division of Urology, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Masashi Honda
- Division of Urology, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Atsushi Takenaka
- Division of Urology, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan
| | - Futoshi Okada
- Division of Pathological Biochemistry, Faculty of Medicine, Tottori University, Tottori 683-8503, Japan.,Chromosome Engineering Research Center, Tottori University, Tottori 683-8503, Japan
| |
Collapse
|
|
18
|
Wang Q, Yao J, Jin Q, Wang X, Zhu H, Huang F, Wang W, Qiang J, Ni Q. LAMP1 expression is associated with poor prognosis in breast cancer. Oncol Lett 2017; 14:4729-4735. [PMID: 29085473 PMCID: PMC5649640 DOI: 10.3892/ol.2017.6757] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/11/2016] [Accepted: 06/15/2017] [Indexed: 01/20/2023] Open
Abstract
Lysosome associated membrane protein-1 (LAMP1) is a heavily glycosylated lysosomal membrane protein, which is able to protect the lysosomal membrane from intracellular proteolysis. LAMP1 has been implicated in cancer development and progression. However, LAMP1 expression in breast cancer (BC) and its relationship with the clinical parameters of BC has not yet been fully investigated. In the present study, LAMP1 expression in BC was characterized by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) on 20 pairs of fresh-frozen BC and corresponding non-cancerous tissues. In addition, tissue microarray immunohistochemistry (TMA-IHC) was conducted on 143 BC and matched non-cancerous tissue samples. The results of RT-qPCR and TMA-IHC demonstrated that LAMP1 expression in BC tissues was significantly higher than in corresponding non-cancerous tissues. Furthermore, LAMP1 protein expression levels were significantly associated with histological grade (P=0.047), estrogen receptor expression (P=0.003), progesterone receptor expression (P=0.002), molecular classification (P=0.022), lymph node metastasis (P=0.033) and tumor-node-metastasis (TNM) stage (P=0.012). Multivariate analysis using Cox regression models and Kaplan-Meier survival curves demonstrated that LAMP1 expression (P=0.037), molecular classification (P=0.017) and TNM stage (P=0.003) were independent prognostic factors for overall survival. The above data suggested that LAMP1 expression is associated with malignant attributes of BC and may serve as a novel prognostic factor for patients with BC.
Collapse
Affiliation(s)
- Qingqing Wang
- Department of General Surgery, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Juan Yao
- Medical School, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China.,Department of Pathology, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Qin Jin
- Department of Pathology, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Xudong Wang
- Surgical Comprehensive Laboratory, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Huijun Zhu
- Department of Pathology, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Fan Huang
- Department of Pathology, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Wei Wang
- Department of Pathology, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Jianfeng Qiang
- Medical School, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| | - Qichao Ni
- Department of General Surgery, The Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China
| |
Collapse
|
|
19
|
Matboli M, Shafei AE, Shehata HH, Nabil N, Hossam N, Azazy AE, El-Tawdi AH, Abdel-Rahman O. Clinical significance of miRNA-autophagy transcript expression in patients with hepatocellular carcinoma. Biomark Med 2017; 11:641-656. [PMID: 28770611 DOI: 10.2217/bmm-2017-0049] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022] Open
Abstract
AIM This study integrates autophagy transcripts miRNAs expression based on bioinformatic analysis followed by clinical validation. METHODOLOGY Cellular jun proto-oncogene mRNA, LAMP2 mRNA, miR-16 and miR-146a level were investigated in the serum and tissue of patients with hepatocellular carcinoma (HCC), chronic hepatitis C and healthy volunteers by quantitative real-time PCR. The prognostic power of this serum RNA panel was explored. RESULTS The expression of serum cellular jun proto-oncogene mRNA, LAMP2 mRNA, miR-16 and miR-146a were positive in 85.1, 94, 97.1 and 84.2% HCC patients, respectively and they were correlated with tissue levels. Our results suggested that the chosen panel is an independent prognostic factor for survival in patients with HCC. CONCLUSION The current work provides four RNA-based biomarker panel for HCC diagnosis and prognosis.
Collapse
Affiliation(s)
- Marwa Matboli
- Oncology Diagnostic Unit, Medical Biochemistry & Molecular biology Department, Faculty of Medicine, Ain Shams University, PO box 11381, Abbassia, Cairo, Egypt
| | - Ayman E Shafei
- Biomedical Research Department, Military Armed Forces College of Medicine
| | - Hanan H Shehata
- Oncology Diagnostic Unit, Medical Biochemistry & Molecular biology Department, Faculty of Medicine, Ain Shams University, PO box 11381, Abbassia, Cairo, Egypt
| | - Nesreen Nabil
- Department of Biochemistry, Faculty of pharmacy, Modern Univesity for Technology & Information, Cairo, Egypt
| | - Nourhan Hossam
- Oncology Diagnostic Unit, Medical Biochemistry & Molecular biology Department, Faculty of Medicine, Ain Shams University, PO box 11381, Abbassia, Cairo, Egypt
| | - Ahmed Em Azazy
- Undergraduate Student, Armed Forces College of Medicine, Cairo, Egypt
| | | | - Omar Abdel-Rahman
- Clinical Oncology Department, Faculty of Medicine, Ain Shams University
| |
Collapse
|
|
20
|
Alessandrini F, Pezzè L, Ciribilli Y. LAMPs: Shedding light on cancer biology. Semin Oncol 2017; 44:239-253. [PMID: 29526252 DOI: 10.1053/j.seminoncol.2017.10.013] [Citation(s) in RCA: 100] [Impact Index Per Article: 12.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/29/2017] [Revised: 10/27/2017] [Accepted: 10/29/2017] [Indexed: 01/09/2023]
Abstract
Lysosomes are important cytoplasmic organelles whose critical functions in cells are increasingly being understood. In particular, despite the long-standing accepted concept about the role of lysosomes as cellular machineries solely assigned to degradation, it has been demonstrated that they play active roles in homeostasis and even in cancer biology. Indeed, it is now well documented that during the process of cellular transformation and cancer progression lysosomes are changing localization, composition, and volume and, through the release of their enzymes, lysosomes can also enhance cancer aggressiveness. LAMPs (lysosome associated membrane proteins) represent a family of glycosylated proteins present predominantly on the membrane of lysosomes whose expression can vary among different tissues, suggesting a separation of functions. In this review we focus on the functions and roles of the different LAMP family members, with a particular emphasis on cancer progression and metastatic spread. LAMP proteins are involved in many different aspects of cell biology and can influence cellular processes such as phagocytosis, autophagy, lipid transport, and aging. Interestingly, for all the five members identified so far (LAMP1, LAMP2, LAMP3, CD68/Macrosialin/LAMP4, and BAD-LAMP/LAMP5), a role in cancer has been suggested. While this is well documented for LAMP1 and LAMP2, the involvement of the other three proteins in cancer progression and aggressiveness has recently been proposed and remains to be elucidated. Here we present different examples about how LAMP proteins can influence and support tumor growth and metastatic spread, emphasizing the impact of each single member of the family.
Collapse
Affiliation(s)
- Federica Alessandrini
- Laboratory of Molecular Cancer Genetics, Centre for Integrative Biology (CIBIO), University of Trento, Povo (TN), Italy
| | - Laura Pezzè
- Laboratory of Molecular Cancer Genetics, Centre for Integrative Biology (CIBIO), University of Trento, Povo (TN), Italy
| | - Yari Ciribilli
- Laboratory of Molecular Cancer Genetics, Centre for Integrative Biology (CIBIO), University of Trento, Povo (TN), Italy.
| |
Collapse
|
|
21
|
Costa J. Glycoconjugates from extracellular vesicles: Structures, functions and emerging potential as cancer biomarkers. Biochim Biophys Acta Rev Cancer 2017; 1868:157-166. [PMID: 28347750 DOI: 10.1016/j.bbcan.2017.03.007] [Citation(s) in RCA: 53] [Impact Index Per Article: 6.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2017] [Revised: 03/20/2017] [Accepted: 03/21/2017] [Indexed: 02/08/2023]
Abstract
Extracellular vesicles (EVs) are released by virtually all cells, carry cellular molecules to the extracellular environment, and may interact with other cells. They are found in body fluids, therefore, constituting useful target sources for the identification of disease biomarkers, for example, in cancer. EVs originate from the plasma membrane or from multivesicular endosomes. They have the same topology as the plasma membrane and are rich in glycoconjugates, displaying specific glycosignatures. Surface glycoconjugates play important roles in EVs biogenesis and in their interaction with other cells. Changes in glycosylation constitute a hallmark of different types of cancer, therefore, the study of glycoconjugates and glycosignatures of EVs appear as promising candidates to identify novel cancer biomarkers and to increase the specificity and sensitivity of the existing clinical biomarkers, many of which are glycosylated.
Collapse
Affiliation(s)
- Julia Costa
- Laboratory of Glycobiology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, 2780-157 Oeiras, Portugal.
| |
Collapse
|
|
22
|
Lu M, Zhu H, Wang X, Zhang D, Xiong L, Zhu J, Mao Y, Qiang J. LAMP1 expression is associated with malignant behaviours and predicts unfavourable prognosis in laryngeal squamous cell carcinoma. Pathology 2016; 48:684-690. [PMID: 27788920 DOI: 10.1016/j.pathol.2016.08.001] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/14/2016] [Revised: 05/30/2016] [Accepted: 08/10/2016] [Indexed: 12/27/2022]
Abstract
Lysosome-associated membrane protein-1 (LAMP1) has been suggested to play complicated roles in cancer development and metastasis. The aim of this study was to explore the expression of LAMP1 in laryngeal squamous cell carcinoma (LSCC) and investigate the relationship between LAMP1 expression and clinicopathological characteristics in LSCC patients. One-step quantitative reverse transcription PCR (qPCR) tests (20 fresh LSCC and non-cancerous tissue samples) and immunohistochemistry (IHC) analyses (137 paraffin-embedded LSCC and non-cancerous tissue samples) were performed to evaluate the LAMP1 expression in both mRNA and protein levels. Results showed that the expression of LAMP1 in LSCC tissues was significantly higher than that in non-cancerous tissues. Furthermore, the expression level of LAMP1 protein was statistically associated with lymph node metastasis and TNM stage. Results of Kaplan-Meier survival and Cox regression analyses revealed that LSCC patients with high LAMP1 cytoplasmic expression (p=0.044), high cytoplasmic+low mesenchymal expression of LAMP1 (H+L) (p=0.015) and histopathological grade (p=0.014) encountered poor overall survival. The data implied that high LAMP1 expression is associated with unfavourable prognosis in LSCC patients, and LAMP1 may be identified as a novel prognostic biomarker.
Collapse
Affiliation(s)
- Meiping Lu
- Department of Otolaryngology-Head and Neck Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Huijun Zhu
- Department of Pathology, Affiliated Hospital of Nantong University, Nantong, China
| | - Xudong Wang
- Department of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong, China
| | - Dawei Zhang
- Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Lin Xiong
- Department of Pathology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China
| | - Jin Zhu
- The Key Laboratory of Cancer Biomarkers, Prevention and Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
| | - Yuan Mao
- Department of Hematology and Oncology, Jiangsu Province Geriatric Hospital, Nanjing, China.
| | - Jianfeng Qiang
- Graduate College, Medical School of Nantong University, Nantong, China.
| |
Collapse
|
|
23
|
Kitahara T, Haraguchi N, Takahashi H, Nishimura J, Hata T, Takemasa I, Mizushima T, Yamamoto H, Doki Y, Mori M. Identification and Characterization of CD107a as a Marker of Low Reactive Oxygen Species in Chemoresistant Cells in Colorectal Cancer. Ann Surg Oncol 2016; 24:1110-1119. [PMID: 27834032 DOI: 10.1245/s10434-016-5671-8] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2015] [Indexed: 11/18/2022]
Abstract
BACKGROUND Reactive oxygen species (ROS) generated by chemoradiotherapy lead to cancer cell death. Although ROS regulation mechanisms play important roles in chemoradioresistance, few markers exist that indicated intracellular ROS status. This study aimed to identify novel cell surface markers that represented intracellular ROS status to characterize cells with low ROS (ROSlow) in colorectal cancer (CRC). METHODS We used ROS indicators and an antibody array with 242 cell surface antibodies to identify markers of ROSlow cells. After validation, we performed immunohistochemical analyses and chemosensitivity assays. We used small interfering RNA to assess the effect of silencing the identified markers. We tested cell differentiation assays with spheroid cell assays. RESULTS CD107a was identified as a common marker of ROSlow cells in several CRC cell lines and clinical specimens. CD107a+/ROSlow cells were enriched in HT29 and DLD1 cultures after treatments with oxaliplatin, 5-fluorouracil, and the irinotecan metabolite SN38. CD107a silencing improved chemosensitivity by increasing ROS production. Immunohistochemistry showed enhanced CD107a surface expression on cells that formed immature cell clusters and on cells located in the invasive fronts of cancer foci. CD107a expression was also enhanced on specimens from patients with poorly differentiated adenocarcinoma who had received neoadjuvant chemotherapy. Cell surface CD107a expression was enhanced on cells that formed colonospheres, but expression diminished during cell differentiation. CONCLUSIONS CD107a was identified as a novel marker of ROSlow cells in CRC. CD107a expression was closely related to chemoresistance and the immature cell phenotype. Anti-CD107a treatments represent a novel approach for targeting chemoresistant cells in CRC.
Collapse
Affiliation(s)
- Tomohiro Kitahara
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Naotsugu Haraguchi
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
| | - Hidekazu Takahashi
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Junichi Nishimura
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Taishi Hata
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Ichiro Takemasa
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Tsunekazu Mizushima
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Hirofumi Yamamoto
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Yuichiro Doki
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| | - Masaki Mori
- Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan
| |
Collapse
|
|
24
|
El-Tawdi AHF, Matboli M, Shehata HH, Tash F, El-Khazragy N, Azazy AESM, Abdel-Rahman O. Evaluation of Circulatory RNA-Based Biomarker Panel in Hepatocellular Carcinoma. Mol Diagn Ther 2016; 20:265-77. [DOI: 10.1007/s40291-016-0200-9] [Citation(s) in RCA: 35] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/21/2022]
|
|
25
|
Piccolo E, Tinari N, D'Addario D, Rossi C, Iacobelli V, La Sorda R, Lattanzio R, D'Egidio M, Di Risio A, Piantelli M, Natali PG, Iacobelli S. Prognostic relevance of LGALS3BP in human colorectal carcinoma. J Transl Med 2015. [PMID: 26219351 PMCID: PMC4518516 DOI: 10.1186/s12967-015-0606-x] [Citation(s) in RCA: 23] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
Background A previous report has shown that LGALS3BP (also known as 90K or Mac-2 BP) has antitumor activity in colorectal cancer (CRC) via suppression of Wnt signalling with a novel mechanism of ISGylation-dependent ubiquitination of β-catenin. The role of LGALS3BP in CRC prognosis was investigated. Methods The role of LGALS3BP on CRC progression and clinical prognosis was analyzed by combining cell cultures, in vitro assays, and immunohistochemistry. Results Silencing of LGALS3BP in HCT-116 human colon cancer cells resulted in enhanced β-catenin expression that was reversed by addition of human recombinant LGALS3BP. Moreover, intra-tumor delivery of LGALS3BP reduced tumor growth of xenografts originating from LGALS3BP-silenced HCT-116 cells. Finally, in a series of 196 CRC patients, LGALS3BP expression in tumor tissue associated with clinical outcome. Patients with high LGALS3BP expression had lower risk of relapse and a longer overall survival time than those with low LGALS3BP expression. Multivariate analyses confirmed LGALS3BP expression status as the only independent prognostic factor of survival. Conclusions These results provide evidence that low expression of LGALS3BP participates in malignant progression of CRC and implicates poor prognosis, highlighting its augmentation as a potential therapeutic approach.
Collapse
Affiliation(s)
- Enza Piccolo
- MediaPharma s.r.l., Via dei Vestini, 31, Chieti, Italy.
| | - Nicola Tinari
- MediaPharma s.r.l., Via dei Vestini, 31, Chieti, Italy. .,Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| | - Domenica D'Addario
- Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| | - Cosmo Rossi
- Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| | | | | | - Rossano Lattanzio
- Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| | - Maurizia D'Egidio
- Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| | | | - Mauro Piantelli
- MediaPharma s.r.l., Via dei Vestini, 31, Chieti, Italy. .,Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| | | | - Stefano Iacobelli
- MediaPharma s.r.l., Via dei Vestini, 31, Chieti, Italy. .,Department of Experimental and Clinical Sciences, "G. D'Annunzio" University and Foundation, Chieti, Italy.
| |
Collapse
|
|
26
|
Lin TW, Chang HT, Chen CH, Chen CH, Lin SW, Hsu TL, Wong CH. Galectin-3 Binding Protein and Galectin-1 Interaction in Breast Cancer Cell Aggregation and Metastasis. J Am Chem Soc 2015; 137:9685-93. [PMID: 26168351 DOI: 10.1021/jacs.5b04744] [Citation(s) in RCA: 49] [Impact Index Per Article: 4.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022]
Abstract
Galectin-3 binding protein (Gal-3BP) is a large hyperglycosylated protein that acts as a ligand for several galectins through glycan-dependent interactions. Gal-3BP can induce galectin-mediated tumor cell aggregation to increase the survival of cancer cells in the bloodstream during the metastatic process. However, the galectin interacting with Gal-3BP and its binding specificity has not been identified and structurally elucidated, mainly due to the limitation of mass spectrometry in glycan sequencing. To understand the role of Gal-3BP, we here used liquid chromatography-mass spectrometry combined with specific exoglycosidase reactions to determine the sequences of N-glycans on Gal-3BP from MCF-7 and MDA-MB-231 cells, especially the sequences with terminal sialylation and fucosylation, and addition of LacNAc repeat structures. The N-glycans from both strains are complex type with terminal α2,3-sialidic acid and core fucose linkages, with additional α1,2- and α1,3 fucose linkages found in MCF-7 cells. Compared with that from MCF-7, the Gal-3BP from MDA-MB-231 cells had fewer tetra-antennary structures, only α1,6-linked core fucoses, and more LacNAc repeat structures; the MDA-MB-231 cells had no surface galectin-3 but used surface galectin-1 for interaction with Gal-3BP to form large oligomers and cell aggregates. This study elucidates the specificity of Gal-3BP interacting with galectin-1 and the role of Gal-3BP in cancer cell aggregation and metastasis.
Collapse
Affiliation(s)
| | - Hui-Tzu Chang
- §Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 112, Taiwan
| | | | | | | | | | | |
Collapse
|
|
27
|
Seol HS, Akiyama Y, Shimada S, Lee HJ, Kim TI, Chun SM, Singh SR, Jang SJ. Epigenetic silencing of microRNA-373 to epithelial-mesenchymal transition in non-small cell lung cancer through IRAK2 and LAMP1 axes. Cancer Lett 2014; 353:232-41. [PMID: 25063738 PMCID: PMC7707239 DOI: 10.1016/j.canlet.2014.07.019] [Citation(s) in RCA: 57] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/23/2014] [Revised: 07/11/2014] [Accepted: 07/14/2014] [Indexed: 12/21/2022]
Abstract
The role of microRNAs (miRNAs) in carcinogenesis as tumor suppressors or oncogenes has been widely reported. Epigenetic change is one of the mechanisms of transcriptional silencing of miRNAs in cancer. To identify lung cancer-related miRNAs that are mediated by histone modification, we conducted microarray analysis in the Calu-6 non-small cell lung cancer (NSCLC) cell line after treatment with suberoylanilide hydroxamic acid (SAHA), a histone deacetylase (HDAC) inhibitor. The expression level of miR-373 was enhanced by SAHA treatment in this cell line by microarray and the following quantitative RT-PCR analyses. Treatment with another HDAC inhibitor, Trichostatin A, restored the levels of miR-373 expression in A549 and Calu-6 cells, while demethylation drug treatment did not. Importantly, miR-373 was found to be down-regulated in NSCLC tissues and cell lines. Transfection of miR-373 into A549 and Calu-6 cells attenuated cell proliferation, migration, and invasion and reduced the expression of mesenchymal markers. Additional microarray analysis of miR-373-transfected cells and computational predictions identified IRAK2 and LAMP1 as targets of miR-373. Knockdown of these two genes showed similar biological effects to those of miR-373 overexpression. In clinical samples, overexpression of IRAK2 correlated with decreased disease-free survival of patients with non-adenocarcinoma. In conclusion, we found that miR-373 is silenced by histone modification in lung cancer cells and identified its function as a tumor suppressor and negative regulator of the mesenchymal phenotype through downstream IRAK2 and LAMP1 target genes.
Collapse
Affiliation(s)
- Hyang Sook Seol
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea; Asan Center for Cancer Genome Discovery, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea
| | - Yoshimitsu Akiyama
- Department of Molecular Oncology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo 113-8519, Japan
| | - Shu Shimada
- Department of Molecular Oncology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo 113-8519, Japan
| | - Hee Jin Lee
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea
| | - Tae Im Kim
- Asan Center for Cancer Genome Discovery, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea
| | - Sung Min Chun
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea; Asan Center for Cancer Genome Discovery, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea
| | - Shree Ram Singh
- Basic Research Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
| | - Se Jin Jang
- Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea; Asan Center for Cancer Genome Discovery, University of Ulsan College of Medicine, Asan Medical Center, Seoul 138-736, South Korea.
| |
Collapse
|
|
28
|
Kim S, Park T, Kon M. Cancer survival classification using integrated data sets and intermediate information. Artif Intell Med 2014; 62:23-31. [DOI: 10.1016/j.artmed.2014.06.003] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2013] [Revised: 04/07/2014] [Accepted: 06/16/2014] [Indexed: 12/11/2022]
|
|
29
|
Marzinke MA, Choi CH, Chen L, Shih IM, Chan DW, Zhang H. Proteomic analysis of temporally stimulated ovarian cancer cells for biomarker discovery. Mol Cell Proteomics 2013; 12:356-68. [PMID: 23172893 PMCID: PMC3567859 DOI: 10.1074/mcp.m112.019521] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2012] [Revised: 11/03/2012] [Indexed: 11/06/2022] Open
Abstract
While ovarian cancer remains the most lethal gynecological malignancy in the United States, there are no biomarkers available that are able to predict therapeutic responses to ovarian malignancies. One major hurdle in the identification of useful biomarkers has been the ability to obtain enough ovarian cancer cells from primary tissues diagnosed in the early stages of serous carcinomas, the most deadly subtype of ovarian tumor. In order to detect ovarian cancer in a state of hyperproliferation, we analyzed the implications of molecular signaling cascades in the ovarian cancer cell line OVCAR3 in a temporal manner, using a mass-spectrometry-based proteomics approach. OVCAR3 cells were treated with EGF(1), and the time course of cell progression was monitored based on Akt phosphorylation and growth dynamics. EGF-stimulated Akt phosphorylation was detected at 12 h post-treatment, but an effect on proliferation was not observed until 48 h post-exposure. Growth-stimulated cellular lysates were analyzed for protein profiles between treatment groups and across time points using iTRAQ labeling and mass spectrometry. The protein response to EGF treatment was identified via iTRAQ analysis in EGF-stimulated lysates relative to vehicle-treated specimens across the treatment time course. Validation studies were performed on one of the differentially regulated proteins, lysosomal-associated membrane protein 1 (LAMP-1), in human tissue lysates and ovarian tumor tissue sections. Further, tissue microarray analysis was performed to demarcate LAMP-1 expression across different stages of epithelial ovarian cancers. These data support the use of this approach for the efficient identification of tissue-based markers in tumor development related to specific signaling pathways. LAMP-1 is a promising biomarker for studies of the progression of EGF-stimulated ovarian cancers and might be useful in predicting treatment responses involving tyrosine kinase inhibitors or EGF receptor monoclonal antibodies.
Collapse
Affiliation(s)
- Mark A. Marzinke
- From the ‡Department of Pathology, Johns Hopkins University, Baltimore, MD 21231
| | - Caitlin H. Choi
- From the ‡Department of Pathology, Johns Hopkins University, Baltimore, MD 21231
| | - Li Chen
- From the ‡Department of Pathology, Johns Hopkins University, Baltimore, MD 21231
| | - Ie-Ming Shih
- From the ‡Department of Pathology, Johns Hopkins University, Baltimore, MD 21231
| | - Daniel W. Chan
- From the ‡Department of Pathology, Johns Hopkins University, Baltimore, MD 21231
| | - Hui Zhang
- From the ‡Department of Pathology, Johns Hopkins University, Baltimore, MD 21231
| |
Collapse
|
|
30
|
Su L, Cao L, Zhou R, Jiang Z, Xiao K, Kong W, Wang H, Deng J, Wen B, Tan F, Zhang Y, Xie L. Identification of novel biomarkers for sepsis prognosis via urinary proteomic analysis using iTRAQ labeling and 2D-LC-MS/MS. PLoS One 2013; 8:e54237. [PMID: 23372690 PMCID: PMC3553154 DOI: 10.1371/journal.pone.0054237] [Citation(s) in RCA: 55] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/03/2012] [Accepted: 12/10/2012] [Indexed: 12/17/2022] Open
Abstract
OBJECTIVES Sepsis is the major cause of death for critically ill patients. Recent progress in proteomics permits a thorough characterization of the mechanisms associated with critical illness. The purpose of this study was to screen potential biomarkers for early prognostic assessment of patients with sepsis. METHODS For the discovery stage, 30 sepsis patients with different prognoses were selected. Urinary proteins were identified using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with LC-MS/MS. Mass spec instrument analysis were performed with Mascot software and the International Protein Index (IPI); bioinformatic analyses were used by the algorithm of set and the Gene Ontology (GO) Database. For the verification stage, the study involved another 54 sepsis-hospitalized patients, with equal numbers of patients in survivor and non-survivor groups based on 28-day survival. Differentially expressed proteins were verified by Western Blot. RESULTS A total of 232 unique proteins were identified. Proteins that were differentially expressed were further analyzed based on the pathophysiology of sepsis and biomathematics. For sepsis prognosis, five proteins were significantly up-regulated: selenium binding protein-1, heparan sulfate proteoglycan-2, alpha-1-B glycoprotein, haptoglobin, and lipocalin; two proteins were significantly down-regulated: lysosome-associated membrane proteins-1 and dipeptidyl peptidase-4. Based on gene ontology clustering, these proteins were associated with the biological processes of lipid homeostasis, cartilage development, iron ion transport, and certain metabolic processes. Urinary LAMP-1 was down-regulated, consistent with the Western Blot validation. CONCLUSION This study provides the proteomic analysis of urine to identify prognostic biomarkers of sepsis. The seven identified proteins provide insight into the mechanism of sepsis. Low urinary LAMP-1 levels may be useful for early prognostic assessment of sepsis. TRIAL REGISTRATION ClinicalTrial.gov NCT01493492.
Collapse
Affiliation(s)
- Longxiang Su
- Department of Respiratory Medicine, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan Province, China
- Medical College, Nankai University, Tianjin, China
- Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing, China
| | - Lichao Cao
- Shenzhen Proteome Engineering Laboratory, BGI Shenzhen, Shenzhen, China
| | - Ruo Zhou
- Shenzhen Proteome Engineering Laboratory, BGI Shenzhen, Shenzhen, China
| | - Zhaoxu Jiang
- Department of Respiratory Medicine, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan Province, China
- Medical College, Nankai University, Tianjin, China
- Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing, China
| | - Kun Xiao
- Department of Respiratory Medicine, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan Province, China
- Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing, China
| | - Weijing Kong
- Department of Pediatrics, First Hospital, Peking University, Beijing, China
| | - Huijuan Wang
- Department of Respiratory Medicine, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan Province, China
- Medical College, Nankai University, Tianjin, China
- Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing, China
| | - Jie Deng
- Department of Respiratory Medicine, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan Province, China
- Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing, China
| | - Bo Wen
- Shenzhen Proteome Engineering Laboratory, BGI Shenzhen, Shenzhen, China
| | - Fengji Tan
- Shenzhen Proteome Engineering Laboratory, BGI Shenzhen, Shenzhen, China
| | - Yong Zhang
- Shenzhen Proteome Engineering Laboratory, BGI Shenzhen, Shenzhen, China
| | - Lixin Xie
- Department of Respiratory Medicine, Hainan Branch of Chinese PLA General Hospital, Sanya, Hainan Province, China
- Department of Respiratory Medicine, Chinese PLA General Hospital, Beijing, China
| |
Collapse
|
|
31
|
Endo H, Muramatsu T, Furuta M, Uzawa N, Pimkhaokham A, Amagasa T, Inazawa J, Kozaki KI. Potential of tumor-suppressive miR-596 targeting LGALS3BP as a therapeutic agent in oral cancer. Carcinogenesis 2012; 34:560-9. [PMID: 23233740 DOI: 10.1093/carcin/bgs376] [Citation(s) in RCA: 56] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/16/2023] Open
Abstract
The incidence and mortality statistics for oral squamous cell carcinoma (OSCC) were 10th and 12th, respectively, in human cancers diagnosed worldwide in 2008. In this study, to identify novel tumor-suppressive microRNAs (TS-miRNAs) and their direct targets in OSCC, we performed methylation-based screening for 43 miRNAs encoded by 46 miRNA genes located within 500 bp downstream of 40 CpG islands and genome-wide gene expression profiling in combination with a prediction database analysis, respectively, in 18 cell lines, resulting in the identification of a novel TS-miRNA miR-596 directly targeting LGALS3BP/Mac-2 BP/90K. DNA hypermethylation of CpG island located 5'-upstream of miR-596 gene was frequently observed in OSCC cell lines (100% of 18 cell lines) and primary OSCC cases (46.2 and 76.3% of 26 Japanese and 38 Thais primary cases, respectively) in a tumor-specific manner. The ectopic transfection of double-stranded RNA (dsRNA) mimicking miR-596 or specific small interfering RNA for LGALS3BP significantly induced growth inhibition and apoptosis in cell lines lacking miR-596 expression or overexpressing LGALS3BP, respectively, in a manner associated with a suppression of ERK1/2 phosphorylation. Moreover, we also mention the effect of dsRNA mimicking miR-596 on the growth of an OSCC cell line in vivo. Our findings define a central role for miR-596 in OSCC and suggest the potential of miR-596 as an anticancer agent for miRNA replacement therapy in OSCC.
Collapse
Affiliation(s)
- Hironori Endo
- Department of Molecular Cytogenetics, Tokyo Medical and Dental University, 1-5-45 Yushima Bunkyo-ku, Tokyo 113-8510, Japan
| | | | | | | | | | | | | | | |
Collapse
|
|
32
|
Huang Y, Goldberg M, Le T, Qiang R, Warner D, Witkowska HE, Liu H, Zhu L, Denbesten P, Li W. Amelogenin exons 8 and 9 encoded peptide enhances leucine rich amelogenin peptide mediated dental pulp repair. Cells Tissues Organs 2012; 196:151-60. [PMID: 22301468 DOI: 10.1159/000331248] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 07/26/2011] [Indexed: 01/09/2023] Open
Abstract
Amelogenins containing exons 8 and 9 are alternatively spliced variants of amelogenin. Some amelogenin spliced variants have been found to promote pulp regeneration following pulp exposure. The function of the amelogenin spliced variants with the exons 8 and 9 remains unknown. In this study, we synthesized recombinant leucine rich amelogenin peptide (LRAP, A-4), LRAP plus exons 8 and 9 peptide (LRAP 8, 9) or exons 8 and 9 peptide (P89), to determine their effects on odontoblasts. In vivo analyses were completed following the insertion of agarose beads containing LRAP or LRAP 8, 9 into exposed cavity preparations of rat molars. After 8, 15 or 30 days' exposure, the pulp tissues were analyzed for changes in histomorphometry and cell proliferation by PCNA stainings. In vitro analyses included the effects of the addition of the recombinant proteins or peptide on cell proliferation, differentiation and adhesion of postnatal human dental pulp cells (DPCs). These studies showed that in vivo LRAP 8, 9 enhanced the reparative dentin formation as compared to LRAP. In vitro LRAP 8, 9 promoted DPC proliferation and differentiation to a greater extent than LRAP. These data suggest that amelogenin exons 8 and 9 may be useful in amelogenin-mediated pulp repair.
Collapse
Affiliation(s)
- Yulei Huang
- Department of Oral Medicine, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, PR China
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
33
|
Kim YS, Jung JA, Kim HJ, Ahn YH, Yoo JS, Oh S, Cho C, Yoo HS, Ko JH. Galectin-3 binding protein promotes cell motility in colon cancer by stimulating the shedding of protein tyrosine phosphatase kappa by proprotein convertase 5. Biochem Biophys Res Commun 2010; 404:96-102. [PMID: 21094132 DOI: 10.1016/j.bbrc.2010.11.071] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/02/2010] [Accepted: 11/16/2010] [Indexed: 01/11/2023]
Abstract
It has previously been reported that shedding of the PTPκ ectodomain drives enhanced motility of colon cancer cells. Herein, we provide mechanism underlying the regulation of PTPκ shedding by galectin-3 binding protein. PTPκ was inarguably scissored by the processed form of proprotein convertase 5 (subtilisin/kexin type 5), and galectin-3 binding protein which is over-produced in colon cancer cells and tissues contributed to increased cancer cell motility by acting as a negative regulator of galectin-3 at the cell surface. The high expression ratio of galectin-3 binding protein to galectin-3 was clinically correlated to lymphatic invasion. These results suggest that galectin-3 binding protein may be a potential therapeutic target for treatment of, at least, colon cancer patients with high expression of galectin-3 binding protein.
Collapse
Affiliation(s)
- Yong-Sam Kim
- Daejeon-KRIBB-FHCRC Research Cooperation Center, KRIBB, Daejeon 305-806, South Korea
| | | | | | | | | | | | | | | | | |
Collapse
|
|
34
|
Liu X, Zhang M, Go VLW, Hu S. Membrane proteomic analysis of pancreatic cancer cells. J Biomed Sci 2010; 17:74. [PMID: 20831833 PMCID: PMC2949717 DOI: 10.1186/1423-0127-17-74] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/05/2010] [Accepted: 09/13/2010] [Indexed: 01/08/2023] Open
Abstract
Background Pancreatic cancer is one of the most aggressive human tumors due to its high potential of local invasion and metastasis. The aim of this study was to characterize the membrane proteomes of pancreatic ductal adenocarcinoma (PDAC) cells of primary and metastatic origins, and to identify potential target proteins related to metastasis of pancreatic cancer. Methods Membrane/membrane-associated proteins were isolated from AsPC-1 and BxPC-3 cells and identified with a proteomic approach based on SDS-PAGE, in-gel tryptic digestion and liquid chromatography with tandem mass spectrometry (LC-MS/MS). X! Tandem was used for database searching against the SwissProt human protein database. Results We identified 221 & 208 proteins from AsPC-1 and BxPC-3 cells, respectively, most of which are membrane or membrane-associated proteins. A hundred and nine proteins were found in both cell lines while the others were present in either AsPC-1 or BxPC-3 cells. Differentially expressed proteins between two cell lines include modulators of cell adhesion, cell motility or tumor invasion as well as metabolic enzymes involved in glycolysis, tricarboxylic acid cycle, or nucleotide/lipid metabolism. Conclusion Membrane proteomes of AsPC-1 (metastatic) and BxPC-3 (primary) cells are remarkably different. The differentially expressed membrane proteins may serve as potential targets for diagnostic and therapeutic interventions.
Collapse
Affiliation(s)
- Xiaojun Liu
- UCLA School of Dentistry & Dental Research Institute, Los Angeles, CA 90095, USA
| | | | | | | |
Collapse
|
|
35
|
Bogoeva VP, Varriale A, John CM, D'Auria S. Human galectin-3 interacts with two anticancer drugs. Proteomics 2010; 10:1946-53. [PMID: 20209510 DOI: 10.1002/pmic.200900581] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
Abstract
Human galectin-3 (hGal-3) is a mammalian lectin involved in regulation of RNA splicing, apoptosis, cell differentiation, and proliferation. Multimerized extracellular hGal-3 is thought to crosslink cells by binding to glycoproteins and glycosylated cancer antigens on the cell surface or extracellular matrix. Fluorescence spectroscopy and circular dichroism were used to study the interaction of hGal-3 with two anticancer agents: bohemine and Zn porphyrin (ZnTPPS(4)). The dissociation constant (k(D)) for binding of bohemine with hGal-3 was k(D) 0.23+/-0.05 microM. The hyperbolic titration curve indicated the presence of a single bohemine binding site. The binding of ZnTPPS(4) to hGal-3 (with and without lactose) is of high affinity having k(D)=0.18-0.20 microM and is not inhibited by lactose, indicating that ZnTPPS(4) and carbohydrate bind different sites. Circular dichroism spectra of the hGal-3 complexes suggested that the binding of the hydrophobic compounds changed the hGal-3 secondary structure. In summary, we show that two compounds with anticancer activity, bohemine and ZnTPPS(4), have high affinity for hGal-3 at a site that is distinct from its carbohydrate site. Since hGal-3 binds to several carbohydrate cancer antigens, the results suggest that it may have utility in the targeted delivery of drugs for cancer.
Collapse
Affiliation(s)
- Vanya P Bogoeva
- Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
| | | | | | | |
Collapse
|
|
36
|
Lee JH, Zhang X, Shin BK, Lee ES, Kim I. Mac-2 binding protein and galectin-3 expression in mucinous tumours of the ovary: an annealing control primer system and immunohistochemical study. Pathology 2009; 41:229-33. [PMID: 19291534 DOI: 10.1080/00313020902756279] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/28/2023]
Abstract
AIM We used a new differential display method, the annealing control primer (ACP) system, to analyse the differentially expressed genes in mucinous ovarian tumours. To verify the corresponding target gene, immunohistochemical staining was performed on various epithelial tumours of the ovary. METHODS AND RESULTS The ACP-based reverse transcriptase-polymerase chain reaction revealed that 21 genes were upregulated in the mucinous ovarian adenoma and 14 genes were upregulated in the mucinous ovarian carcinoma. Among them, we selected one upregulated gene, the Mac-2 binding protein (Mac-2 BP), and verified the expression of the Mac-2 BP and its ligand, galectin-3, in a variety of epithelial ovarian tumours by immunohistochemistry. Positive expression of the Mac-2 BP was significantly higher in the mucinous ovarian tumours compared to the other epithelial tumours. Mac-2 BP expression was significantly increased in the borderline and malignant tumours compared to the benign tumours. Galectin-3 expression was more frequent in clear cell carcinomas, serous tumours and mucinous tumours than in endometrioid and transitional tumours. However, there were no differences in galectin-3 expression in comparisons among benign, borderline and malignant mucinous and serous tumours. CONCLUSION These data indicate that the Mac-2 BP may play a role in the development and progression of mucinous ovarian tumours.
Collapse
Affiliation(s)
- Ju-Han Lee
- Department of Pathology, Medical College, Korea University, Seoul, Korea
| | | | | | | | | |
Collapse
|
|
37
|
Ulmer TA, Keeler V, André S, Gabius HJ, Loh L, Laferté S. The tumor-associated antigen 90K/Mac-2-binding protein secreted by human colon carcinoma cells enhances extracellular levels of promatrilysin and is a novel substrate of matrix metalloproteinases-2, -7 (matrilysin) and -9: Implications of proteolytic cleavage. Biochim Biophys Acta Gen Subj 2009; 1800:336-43. [PMID: 19665518 DOI: 10.1016/j.bbagen.2009.07.030] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/20/2009] [Revised: 07/24/2009] [Accepted: 07/28/2009] [Indexed: 01/07/2023]
Abstract
BACKGROUND The tumor-associated antigen 90K (TAA90K)/Mac-2-binding protein is expressed at elevated level in cancerous tissues and associated with poor prognosis. Since TAA90K has been implicated in the restructuring of the extracellular matrix, we examined the functional relationship between colon cancer cell-derived TAA90K and the matrix metalloproteinase (MMP) promatrilysin (proMMP-7), and also tested whether TAA90K is a novel substrate for MMPs-2, -7 and -9. METHODS The effect of TAA90K on proMMP-7 levels in HT-29 conditioned media was quantified by enzyme-linked immunosorbent assays. Binding of TAA90K to MMPs, extracellular matrix proteins and galectin-3 was measured by solid-phase binding assays. Proteolytic cleavage of TAA90K by MMPs was documented by SDS-PAGE and protein sequencing analysis. RESULTS TAA90K enhanced extracellular levels of proMMP-7 in HT-29 cells. In addition, TAA90K was cleaved by MMPs-2, -7 and -9. MMP-7-mediated cleavage of TAA90K did not affect its binding to MMP-7, laminin-1, collagen IV and galectin-3 but reduced its interaction with fibronectin and laminin-10, and lowered the levels of proMMP-7 in the HT-29 medium. CONCLUSION TAA90K is a novel substrate for MMPs-2, -7 and -9 and modulates proMMP-7 levels in colon cancer cells. GENERAL SIGNIFICANCE Proteolytic cleavage of TAA90K may have functional implications in colon cancer.
Collapse
Affiliation(s)
- Tricia A Ulmer
- Department of Biochemistry, University of Saskatchewan, 107 Wiggins Road, Room A3, Saskatoon, Saskatchewan, Canada S7N 5E5
| | | | | | | | | | | |
Collapse
|
|
38
|
Torlakovic EE, Keeler V, Wang C, Lim HJ, Lining LA, Laferté S. Cyclophilin C-associated protein (CyCAP) knock-out mice spontaneously develop colonic mucosal hyperplasia and exaggerated tumorigenesis after treatment with carcinogen azoxymethane. BMC Cancer 2009; 9:251. [PMID: 19627619 PMCID: PMC2724547 DOI: 10.1186/1471-2407-9-251] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/22/2008] [Accepted: 07/24/2009] [Indexed: 02/07/2023] Open
Abstract
BACKGROUND The discovery of a "serrated neoplasia pathway" has highlighted the role of hyperplastic lesions of the colon as the significant precursor of colorectal adenocarcinoma. In mice, hyperplasia of the colonic mucosa is a regular phenomenon after a challenge with colonic carcinogens indicating that mucosal hyperproliferation and thickening, even without cytological dysplasia, represents an early pre-malignant change. Cyclophilin C-associated protein (CyCAP) has been described to down-modulate endotoxin signaling in colorectal murine mucosa and is a murine orthologue of the tumor-associated antigen 90 K (TAA90K)/mac-2-binding protein. METHODS Female Balb/c wild-type (WT) and CyCAP knock-out (KO) mice (6-8 weeks old) were administered 2 or 6 weekly subcutaneous injections of azoxymethane. The animals were evaluated post-injection at six weeks for aberrant crypt foci (ACF) study and at five months for colon tumor measurement. The thickness of the colon crypts was measured in microns and the number of colonocytes per crypt was also determined in well-oriented crypts. Morphometric analyses of the colon mucosa were also performed in untreated 6-8 weeks old KO and WT animals. Formalin-fixed/paraffin-embedded colon sections were also studied by immunohistochemistry to determine the Ki-67 proliferation fraction of the colon mucosa, beta-catenin cellular localization, cyclin D1, c-myc, and lysozyme in Paneth cells. RESULTS Cyclophilin C-associated protein (CyCAP)-/- mice, spontaneously developed colonic mucosal hyperplasia early in life compared to wild-type mice (WT) (p < 0.0001, T-test) and crypts of colonic mucosa of the (CyCAP)-/- mice show higher proliferation rate (p = 0.039, Mann-Whitney Test) and larger number of cyclin D1-positive cells (p < 0.0001, Mann-Whitney Test). Proliferation fraction and cyclin D1 expression showed positive linear association (p = 0.019, Linear-by-Linear Association). The hyperplasia was even more pronounced in CyCAP-/- mice than in WT after challenge with azoxymethane (p = 0.005, T-test). The length of the crypts (r = 0.723, p = 0.018, Spearman Correlation) and the number of colonocytes per crypt (r = 0.863, p = 0.001, Spearman Correlation) in non-tumorous areas were positively associated with azoxymethane-induced number of tumors. CyCAP-/- developed larger numbers of tumors than WT animals (p = 0.003, T-Test) as well as overall larger tumor mass (p = 0.016, T-Test). Membranous beta-catenin was focally overexpressed in KO mice including proliferative zone of the crypts. CONCLUSION CyCAP-/- represent the first described model of spontaneous colonic mucosal hyperplasia. We conclude that CyCAP-deficient mice spontaneously and after challenge with carcinogen develop significantly more colorectal mucosal hyperplasia, an early stage in murine colonic carcinogenesis.
Collapse
Affiliation(s)
- Emina Emilia Torlakovic
- Department of Pathology, Royal University Hospital, College of Medicine, University of Saskatchewan, Saskatoon, Canada.
| | | | | | | | | | | |
Collapse
|
|
39
|
Fortunato F, Bürgers H, Bergmann F, Rieger P, Büchler MW, Kroemer G, Werner J. Impaired autolysosome formation correlates with Lamp-2 depletion: role of apoptosis, autophagy, and necrosis in pancreatitis. Gastroenterology 2009; 137:350-60, 360.e1-5. [PMID: 19362087 DOI: 10.1053/j.gastro.2009.04.003] [Citation(s) in RCA: 179] [Impact Index Per Article: 11.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/23/2008] [Revised: 03/13/2009] [Accepted: 04/02/2009] [Indexed: 12/17/2022]
Abstract
BACKGROUND & AIMS Acute pancreatitis constitutes a life-threatening condition in which pancreatic acinar cells undergo massive cell death. We investigated the incidence of apoptosis, autophagy, and necrosis affecting acinar cells in the early onset of acute pancreatitis induced by chronic alcohol feeding and acute endotoxemia. METHODS Rats were fed either an ethanol-containing or a control diet over 14 weeks and killed 3 or 24 hours after a single lipopolysaccharide injection. Apoptosis, necrosis, and autophagy of pancreatic acinar cells were assessed by histology, electron microscopy, immunofluorescence, and biochemical methods. RESULTS The combination of alcohol exposure and endotoxemia resulted in the depletion of several lysosomal proteins including lysosomal-associated membrane protein-2 (Lamp-2), a protein that is required for the proper fusion of autophagosomes with lysosomes. Accordingly, Lamp-2 depletion correlated with the accumulation of autophagosomes and a relative paucity of autolysosomes, reduced adenosine-5'-triphosphate levels, and a switch from apoptotic to necrotic cell death. This switch to necrosis was accompanied by reduced caspase activation and the nuclear release of the proinflammatory factor high mobility group box 1. Importantly, human patients with alcoholic pancreatitis also exhibited local Lamp-2 depletion, which points to a crucial role for Lamp-2 and autophagy in pancreatic acinar cell death. CONCLUSIONS Our data suggest that acinar cell vacuolization in pancreatitis is mediated by an endotoxemia-induced inhibition of the late stage of autophagy. The combination of alcohol and endotoxemia attenuated apoptosis response yet enhanced acinar cell necrosis. The depletion of lysosomal proteins plays a critical role in the early onset of acute pancreatitis.
Collapse
Affiliation(s)
- Franco Fortunato
- Department of General, Visceral, and Transplantation Surgery, University Hospital Heidelberg, Germany.
| | | | | | | | | | | | | |
Collapse
|
|
40
|
Tan LB, Chen KT, Yuan YC, Liao PC, Guo HR. Identification of urine PLK2 as a marker of bladder tumors by proteomic analysis. World J Urol 2009; 28:117-22. [PMID: 19506885 DOI: 10.1007/s00345-009-0432-y] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2008] [Accepted: 05/20/2009] [Indexed: 11/29/2022] Open
|
|
41
|
Park YP, Choi SC, Kim BY, Kim JT, Song EY, Kang SH, Yoon DY, Paik SG, Kim KD, Kim JW, Lee HG. Induction of Mac-2BP by nerve growth factor is regulated by the PI3K/Akt/NF-κB-dependent pathway in the HEK293 cell line. BMB Rep 2008; 41:784-9. [DOI: 10.5483/bmbrep.2008.41.11.784] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
|
|
42
|
Soltermann A, Ossola R, Kilgus-Hawelski S, von Eckardstein A, Suter T, Aebersold R, Moch H. N-glycoprotein profiling of lung adenocarcinoma pleural effusions by shotgun proteomics. Cancer 2008; 114:124-33. [PMID: 18327805 DOI: 10.1002/cncr.23349] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
BACKGROUND Malignant pleural effusion of advanced lung adenocarcinoma may be a valid source for detection of biomarkers, such as N-glycosylated proteins (N-GP), because tumor cells grow during weeks in this liquid. The authors aimed for creation of N-GP effusion profiles from routine cytology specimens to detect relevant biomarkers. METHODS Hundred microliters of malignant pleural effusions of 5 patients with lung adenocarcinoma and 5 nonmalignant controls were used for triplicate N-GP capture by solid-phase extraction. After trypsin digest and PNGase F release, a liquid chromatography separation connected online to a tandem mass spectrometer was performed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). RESULTS In the total of 10 samples, 170 and 278 nonredundant proteins were detected with probabilities of >or=.9 and >or=.5, respectively. The specificity for the N-glycomotif was 88% at P >or= .9. Penetration into the moderate to low protein concentration range (microg-ng/mL) occurred, and several proteins associated with tumor progression or metastasis were identified, including CA-125, CD44, CD166, lysosome-associated membrane glycoprotein 2 (LAMP-2), multimerin 2, and periostin. MS identifications were correlated with the corresponding immunoreactivity in either effusion fluid or tumor tissue. CONCLUSIONS In conclusion, reduction of sample complexity by N-GP capturing allows detection of proteins in the mug to ng/mL range. Pleural effusion is a useful source for biomarker research in lung cancer.
Collapse
Affiliation(s)
- Alex Soltermann
- Institute for Surgical Pathology, University Hospital Zurich, Zurich Switzerland.
| | | | | | | | | | | | | |
Collapse
|
|
43
|
Aspinall-O'Dea M, Costello E. The pancreatic cancer proteome - recent advances and future promise. Proteomics Clin Appl 2007; 1:1066-79. [DOI: 10.1002/prca.200700144] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
|
|
44
|
Park YP, Choi SC, Kim JH, Song EY, Kim JW, Yoon DY, Yeom YI, Lim JS, Kim JW, Paik SG, Lee HG. Up-regulation of Mac-2 binding protein by hTERT in gastric cancer. Int J Cancer 2007; 120:813-20. [PMID: 17131321 DOI: 10.1002/ijc.22369] [Citation(s) in RCA: 44] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/11/2023]
Abstract
Mac-2 binding protein (Mac-2BP) is a secreted tumor antigen that is elevated in many cancers and implicated in tumor metastasis, as well as cell adhesion and immune functions. We focused on the human telomerase reverse transcriptase (hTERT) induced Mac-2BP expression and the relationship between Mac-2BP expression and the progression of gastric cancer. A cDNA expression array analysis was performed on the telomerase-negative cell line, SW13, which was engineered to overexpress hTERT when compared with the parental SW13 cell. hTERT-induced Mac-2BP expression was confirmed via RT-PCR and Northern blotting. ELISA and flow cytometric analyses revealed that Mac-2BP protein was increased by 2- to 4-fold in hTERT-overexpressing cells compared with the mock control. Mac-2BP expression was significantly reduced when the overexpressed hTERT was neutralized by the introduction of hTERT-specific siRNA. These results suggest that Mac-2BP expression is modulated by hTERT. Mac-2BP levels in both gastric cancer cells and tumor tissues were determined via Northern blot analysis and immunohistochemistry. Mac-2BP protein was highly expressed in most gastric cancer cell lines, and gastric tumor tissues were stained more densely than normal tissues. The intracellular and secreted Mac-2BP levels were also evaluated via ELISA, indicating that Mac-2BP was expressed and secreted more abundantly in gastric cancer patients than in healthy donors. The elevated serum Mac-2BP level in gastric tumor patients was also significantly associated with distant metastasis (p = 0.05) and higher tumor stage (p = 0.04). Our findings suggest that Mac-2BP is induced by hTERT, and that it may prove to be a useful prognostic marker for the detection of malignant progression of metastatic stomach cancers.
Collapse
|
|
45
|
Ulmer TA, Keeler V, Loh L, Chibbar R, Torlakovic E, André S, Gabius HJ, Laferté S. Tumor-associated antigen 90K/Mac-2-binding protein: possible role in colon cancer. J Cell Biochem 2006; 98:1351-66. [PMID: 16518858 DOI: 10.1002/jcb.20784] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/01/2023]
Abstract
The tumor-associated antigen 90K (TAA90K)/Mac-2-binding protein implicated in cancer progression and metastasis is modified by beta1-6 branched N-linked oligosaccharides in colon cancer cells, glycans shown to contribute to cancer metastasis. To elucidate the role of TAA90K in colon cancer, we examined its expression and function in human colon tumors and colon carcinoma cell lines. Immunohistochemical analyses of colon tumors revealed elevated expression of TAA90K in all samples analyzed compared to normal colon. To examine the function of TAA90K in colon cancer, we carried out protein and cell binding assays using TAA90K-His purified from HT-29 cells colon carcinoma cells infected with recombinant vaccinia virus expressing TAA90K containing a C-terminal poly-histidine tag. TAA90K-His bound to fibronectin, collagen IV, laminins-1, -5, and -10 and galectin-3 (Mac-2) but poorly to collagen I and galectin-1. As expected, binding of TAA90K to galectin-3 was dependent on carbohydrate since it was inhibitable by lactose and asiolofetuin, and a TAA90K-His glycoform purified from HT-29 cells treated with the glycosylation inhibitor 1-deoxymannojirimycin bound poorly to galectin-3. Unlike TAA90K isolated from other cell types, TAA90K-His isolated from colon cancer cells failed to mediate adhesion of colon cancer and normal cell lines, possibly due to cell-type specific glycosylation of TAA90K-His and/or its putative cellular receptor. However, at low concentrations, TAA90K-His enhanced galectin-3-mediated HT-29 cell adhesion while at high concentrations, it inhibited cell adhesion. Thus, a possible mechanism by which TAA90K may contribute to colon cancer progression is by modulating tumor cell adhesion to extracellular proteins, including galectin-3.
Collapse
Affiliation(s)
- Tricia A Ulmer
- Department of Biochemistry, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5, Canada
| | | | | | | | | | | | | | | |
Collapse
|
|
46
|
Modulating expression of LAMPs and ABH histo-blood group antigens in normal and neoplastic human skin. Open Med (Wars) 2006. [DOI: 10.2478/s11536-006-0012-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
AbstractAlthough the precise biological role of lysosomal membrane-associated glycoproteins (LAMPs) and ABH histo-blood group antigens (HBGAs) remains somewhat unclear, they are thought to be related to cell differentiation, cellular adhesion, and tumorigenesis. Here, we present the first comparative immunohistochemical study of both LAMPs and HBGAs in normal and neoplastic skin. Their localization is compared to that of high molecular weight cytokeratin and cytokeratin MNF 116. LAMPs and HBGA were differentially expressed in the normal stratified squamous epithelium, suggesting that they are involved in the initial steps of the differentiation process, whereas HBGAs are characteristic of terminal keratinocyte differentiation. No change in the reactivity for HBGA was detected in the stratified epithelium overlying squamous cell or basal cell carcinomas, whereas a considerable loss of LAMPs was detected. LAMPs were overexpressed in tumor cells, whereas HBGAs were lost in tumor zones of basocellular carcinomas. In spinocellular carcinomas, HBGAs were detected in tumor keratinocytes and in keratin pearls. These results provide new evidence for the differential expression of LAMPs and HBGAs in the normal stratified squamous epithelium, as well as the presence of a modulating reactivity in basocellular and spinocellular carcinomas, suggesting that these glycoproteins are involved in differentiation and tumorigenesis of human skin.
Collapse
|
|
47
|
Liu MP, Guo XZ, Xu JH, Wang D, Li HY, Cui ZM, Zhao JJ, Ren LN. New tumor-associated antigen SC6 in pancreatic cancer. World J Gastroenterol 2006; 11:7671-5. [PMID: 16437697 PMCID: PMC4727239 DOI: 10.3748/wjg.v11.i48.7671] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM To examine the concentration of a new antigen SC6 (SC6-Ag) recognized by monoclonal antibody (MAb) in patients with pancreatic cancer and other malignant or benign diseases and to understand whether SC6-Ag has any clinical significance in distinguishing pancreatic cancer from other gastrointestinal diseases. METHODS Six hundred and ninety-five serum specimens obtained from 115 patients with pancreatic cancer, 154 patients with digestive cancer and 95 patients with non-digestive cancer were used and classified in this study. Serum specimens obtained from 140 patients with benign digestive disease and 89 patients with non-benign digestive disease served as controls. Ascites was tapped from 16 pancreatic cancer patients, 19 hepatic cancer patients, 16 colonic cancer patients, 10 gastric cancer and 6 severe necrotic pancreatitis patients. The samples were quantitated by solid-phase radioimmunoassay. The cut-off values (CV) of 41, 80, and 118 U/mL were used. RESULTS The average intra- and interassay CV detected by immunoradiometric assay of SC6-Ag was 5.4% and 8.7%, respectively. The sensitivity and specificity were 73.0% and 90.9% respectively. The levels in most malignant and benign cases were within the normal upper limit. Among the 16 pancreatic cancer cases, the concentration of SC6-Ag in ascites was over the normal range in 93.8% patients. There was no significant difference in the concentration of SC6-Ag. Decreased expression of SC6-Ag in sera was significantly related to tumor differentiation. The concentration of SC6-Ag was higher in patients before surgery than after surgery. The specificity of SC6-Ag and CA19-9 was significantly higher than that of ultrasound and computer tomography (CT) in pancreatic cancer patients. Higher positive predictive values were indicated in 92.3% SC6-Ag and 88.5% CA19-9, but lower in 73.8% ultrasound and 76.2% CT. CONCLUSION The combined test of SC6-Ag and CA19-9 may improve the diagnostic rate of primary cancer. The detection of SC6-Ag is valuable in the diagnosis of pancreatic cancer before and after surgery.
Collapse
Affiliation(s)
- Min-Pei Liu
- Department of Experimental Medicine, Northern Hospital, No. 83, Wenhua Road, Shenhe District, Shenyang 110016, Liaoning Province, China.
| | | | | | | | | | | | | | | |
Collapse
|
|
48
|
Tompkins K, George A, Veis A. Characterization of a mouse amelogenin [A-4]/M59 cell surface receptor. Bone 2006; 38:172-80. [PMID: 16214432 DOI: 10.1016/j.bone.2005.08.013] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 04/29/2005] [Revised: 08/03/2005] [Accepted: 08/03/2005] [Indexed: 11/28/2022]
Abstract
Amelogenin proteins comprise up to 90% of the organic matrix of developing enamel in the vertebrate tooth. Alternative splicing of mouse amelogenin pre-mRNA leads to the production of more than 14 protein isoforms, the functions of which are not totally understood. The smaller splice products, [A + 4] or M73 and [A - 4] or M59, have been shown to act differently as signaling molecules affecting odontogenic and other cell types. The mechanisms of these signaling processes, beginning with receptor identification, are not well understood. Utilizing radiolabeled [A - 4], we show here that 3H[A - 4] binds in a saturable fashion to the cell surface of C2C12 mouse fetal myoblasts at 4 degrees C, and not only binds at the surface but is internalized at 37 degrees C. "Far Western" immunohistochemistry performed on sections of E18 mouse incisors and molars with biotin-labeled [A - 4] as the primary ligand demonstrates [A - 4]-biotin binding to polarizing ameloblasts and odontoblasts, cells of the dental follicle, and along the stratum intermedium. Using [A - 4] affinity column chromatography and [A - 4]-biotin label transfer reaction, we have identified a 95 kDa C2C12 cell surface protein which bound [A - 4]. Utilizing Tandem MS (MS/MS) sequencing, we report the novel finding of the 95 kDa murine transmembrane protein, LAMP-1, originally identified as a lysosomal membrane protein that is also found at the cell surface, as an [A - 4] cell binding protein.
Collapse
Affiliation(s)
- Kevin Tompkins
- Department of Cell and Molecular Biology, Northwestern University Feinberg School of Medicine, 303 E. Chicago Ave Ward-13-100 Chicago, IL 60611, USA
| | | | | |
Collapse
|
|
49
|
Bagshaw RD, Mahuran DJ, Callahan JW. Lysosomal membrane proteomics and biogenesis of lysosomes. Mol Neurobiol 2005; 32:27-41. [PMID: 16077181 DOI: 10.1385/mn:32:1:027] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/16/2004] [Accepted: 10/14/2004] [Indexed: 12/30/2022]
Abstract
This review focuses on events involved in the biogenesis of the lysosome. This organelle contains a diverse array of soluble, luminal proteins capable of digesting all the macromolecules in the cell. Altered function of lysosomes or its constituent enzymes has been implicated in a host of human pathologies, including storage diseases, cancer, and infectious and neurodegenerative diseases. Luminal enzymes are well-characterized, and aspects of how they are incorporated into lysosomes are known. However, little is known about the composition of the membrane surrounding the organelle or how the membrane is assembled. Our starting point to study lysosome biogenesis is to define the composition of the membrane by the use of proven methods for purification of lysosomes to near homogeneity and then to characterize membrane-associated and integral lysosomal membrane proteins. This has been achieved using advanced proteomics (electrophoretic or chromatographic separations of proteins followed by time-of-flight mass spectrometric identification of peptide sequences). To date, we have identified 55 proteins in the membrane-associated fraction and 215 proteins in the integral membrane. By applying these methods to mouse models of lysosome dysgenesis (such as BEIGE, Pale Ear, PEARL) that are related to human diseases such as Chediak-Higashi and Hermansky-Pudlak syndromes, it may be possible to define the membrane protein composition of lysosomes in each of these mutants and to determine how they differ from normal. Identifying proteins affected in the respective mutants may provide hints about how they are targeted to the lysosomal membrane and how failure to target them leads to disease; these features are pivotal to understanding lysosome biogenesis and have the potential to implicate lysosomes in a broad range of human pathologies.
Collapse
Affiliation(s)
- Richard D Bagshaw
- Research Institute, The Hospital for Sick Children, University of Toronto, Toronto, Canada
| | | | | |
Collapse
|
|
50
|
Wu CC, Chien KY, Tsang NM, Chang KP, Hao SP, Tsao CH, Chang YS, Yu JS. Cancer cell-secreted proteomes as a basis for searching potential tumor markers: nasopharyngeal carcinoma as a model. Proteomics 2005; 5:3173-82. [PMID: 16035111 DOI: 10.1002/pmic.200401133] [Citation(s) in RCA: 88] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Abstract
Nasopharyngeal carcinoma (NPC) is commonly diagnosed late due to its deep location and vague symptoms. To identify biomarkers for early NPC diagnosis, secreted proteomes of two NPC cell lines were analyzed. Proteins in the NPC cell-line cultured media were systematically identified by SDS-PAGE combined with MALDI-TOF MS. Twenty-three proteins were found in cultured media from both NPC cell lines. Among them, fibronectin, Mac-2 binding protein (Mac-2 BP), and plasminogen activator inhibitor 1 (PAI-1) were further confirmed by Western blot analysis. These three proteins were highly expressed in NPC biopsies, but weakly or not expressed in normal nasopharyngeal tissues. The serum levels of the three proteins were significantly higher in NPC patients (n = 46) than in normal controls (n = 47) (p < 0.01). NPC nude mice model (n = 9) also showed elevated levels of serum Mac-2 BP and PAI-1 compared with tumor-free mice (n = 9) (p < 0.01). Systematic analysis of cancer cell-secreted proteomes combined with animal tumor models can be a feasible, convenient strategy for searching multiple potential tumor markers. Furthermore, our work shows that fibronectin, Mac-2 BP, and PAI-1 may be potential markers for diagnosis of NPC.
Collapse
Affiliation(s)
- Chih-Ching Wu
- Department of Cell and Molecular Biology, Chang Gung University, Tao-Yuan, Taiwan, Republic of China
| | | | | | | | | | | | | | | |
Collapse
|