|
1
|
Gupta M, Nimesh H, Bilgrami AL, Sarwat M. A Saffron-based Polyherbal Formulation DuK Prevents Hepatocellular Carcinoma in Male Wistar Rats. Curr Cancer Drug Targets 2025; 25:335-344. [PMID: 37594101 DOI: 10.2174/1568009623666230818115905] [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: 05/15/2023] [Revised: 06/20/2023] [Accepted: 07/20/2023] [Indexed: 08/19/2023]
Abstract
BACKGROUND Duk is a well-established traditional drug that has been used since time immemorial by Indian practitioners to cure various human ailments. OBJECTIVE The purpose of this study was to explore the anti-cancer activity and the possible mechanism of Duk against diethylnitrosamine (DEN)-initiated hepatocarcinogenesis. METHODS We administered Duk at 3 doses, viz., 75, 150, and 300 mg/kg/day, 2 weeks before the DEN and continued it for 16 weeks. After 1 week of DEN recovery, 2-aminoacetylflourine (2- AAF) was administered to promote hepatocarcinogenesis. RESULTS We found that Duk significantly reduced the DEN and 2-AAF induced phenotypical changes in rats and restored the levels of liver function markers. Furthermore, Duk counteracted the oxidative stress induced by carcinogens as observed by restoration in the levels of superoxide dismutase (SOD) and catalase (CAT). Duk significantly diminished the levels of malondialdehyde (MDA) in a dose dependent manner and restored the liver microarchitecture as assessed by histopathological studies. The results of immunohistochemical staining showed that Duk inhibited the DEN-induced decrease in the number of cells positive for Bid and Caspase-9. It also reduces the number of cells positive for Cyclin D. CONCLUSION Duk significantly protects rat liver from hepatocarcinogenesis by regulating oxidative damage and restoring liver function markers. The chemopreventive effect of Duk might be through the induction of apoptosis.
Collapse
Affiliation(s)
- Meenakshi Gupta
- Amity Institute of Pharmacy, Amity University, Noida, Uttar Pradesh, India
| | - Hemlata Nimesh
- Amity Institute of Pharmacy, Amity University, Noida, Uttar Pradesh, India
| | - Anwar L Bilgrami
- Deanship of Scientific Research, King Abdulaziz University, Jeddah, Saudi Arabia
| | - Maryam Sarwat
- Amity Institute of Pharmacy, Amity University, Noida, Uttar Pradesh, India
| |
Collapse
|
|
2
|
Shaglouf LHF, Ranjpour M, Wajid S, Tandon R, Vasudevan KR, Jain SK. Elevated expression of ISY1, APOA-1, SYNE1, MTG1, and MMP10 at HCC initiation: HCC specific protein network involving interactions of key regulators of lipid metabolism, EGFR signaling, MAPK, and splicing pathways. PROTOPLASMA 2023; 260:651-662. [PMID: 35962262 DOI: 10.1007/s00709-022-01796-5] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/26/2022] [Accepted: 07/14/2022] [Indexed: 06/15/2023]
Abstract
Identification of molecular regulators of hepatocellular carcinoma (HCC) initiation and progression is not well understood. We chemically induced HCC in male Wistar rats by administration of diethyl nitrosamine (DEN) and 2-acetylaminofluorene (2-AFF). Using 2D-electrophoresis and MALDI-TOF-MS/MS analyses, we characterized differentially expressed proteins in liver tissues at early stage of HCC progression. Using RT-PCR analysis, we quantified the mRNA expression of the characterized proteins and validated the transcript expression with tumor tissues of clinically confirmed HCC patients. Using bioinformatic tools, we analyzed a network among the introduced proteins that identified their interacting partners and analyzed the molecular mechanisms associated with signaling pathways during HCC progression. We characterized a protein, namely, pre-mRNA splicing factor 1 homolog (ISY1), which is upregulated at both transcriptome and proteome levels at HCC initiation, progression, and tumor stages. We analyzed the interacting partners of ISY1, namely, APOA-1, SYNE1, MMP10, and MTG1. Real-time PCR analysis confirmed elevated expression of APOA-1 mRNA at HCC initiation, progression, and tumor stages in animals undergoing tumorigenesis. The mRNA expression of the interacting partners was validated with tumor tissues of clinically confirmed liver cancer patients; the analysis revealed significant elevation in expression of transcripts. The transcriptome and proteome analyses complement each other and dysregulation in mRNA and protein expression of these regulators may play critical role in HCC initiation and progression.
Collapse
Affiliation(s)
- Laila H Faraj Shaglouf
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, India
| | - Maryam Ranjpour
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, India.
| | - Saima Wajid
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, India
| | - Rakesh Tandon
- Institute of Gastroenterology, PSRI Hospital, New Delhi, India
| | | | - Swatantra Kumar Jain
- Department of Medical Biochemistry, HIMSR, Jamia Hamdard, New Delhi, 110062, India
| |
Collapse
|
|
3
|
Sakr MA, Al-Azzawi MA, Anis A, Abd El-Aziz AA, Ebeid ME, Shokeer MA, fayed A. The correlation between P53 and COX-2 expression and the pathological alteration in hepatocellular carcinoma. EGYPTIAN JOURNAL OF MEDICAL HUMAN GENETICS 2022. [DOI: 10.1186/s43042-022-00230-y] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/10/2023] Open
Abstract
Abstract
Background
Hepatocellular carcinoma (HCC) is among the highest life-threatening malignancies. On both a molecular and histological level, HCC is a highly heterogeneous malignancy. This study was aimed to study the correlation between the molecular expression of some molecular biomarkers (P53 and Cox-2) and the histopathological alterations in the chemically induced HCC by Diethylnitrosamine (DEN) in Adult female Rats. The liver tumor induction was done by injection of DEN intraperitoneally one, two and three times/week for 2 months by the dose of 50 mg/kg Bw. The histopathological analysis was done and expression level of P53 and cox-2 was detected by quantitative polymerase chain reaction (qRT-PCR) at the end of the experiment.
Results
In this study, Grossly, livers of the groups administered with DEN showed multiple grayish-white macronodules on the outer surface which is dose dependent. Histopathologically, DEN induce multifocal micronodules of hepatocellular carcinoma which characterized by nuclear atypia, clear cell, mitotic figures and necrosis of hepatocytes. P53 mRNA expression to GAPDH, revealed that, there was a statistically significant decrease in HCC groups compared to healthy control group, while Cox-2 mRNA expression was significantly increased in HCC groups than healthy control group.
Conclusions
HCC staging can be achieved by detection the expression of P53, and Cox-2 as molecular markers as it considers noninvasive, rapid and easy method than the histopathological analysis. Finally, Cox-2 could be a therapeutic candidate for HCC due to the higher expression of Cox-2 in HCC lesions.
Collapse
|
|
4
|
Hora S, Asad M, Jain SK, Katare DP. Identification of potential targets with high centrality indicated by diethylnitrosamine + thioacetamide-induced hepatocellular carcinoma model. J Cancer Res Ther 2021; 17:1081-1092. [PMID: 34528568 DOI: 10.4103/jcrt.jcrt_948_20] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/04/2022]
Abstract
Background and Aim Hepatocellular carcinoma (HCC), a primary liver malignancy, represents a continuous challenge to clinicians as it is a leading cause of death due to cancer widely. Early detection is the only hope to cure patients from this deadly disease or possibly increase life expectancy. Mouse models are most acceptable studies as they have ability to manipulate their genome and transcriptome to evaluate mechanistic changes. In addition, system biology can improvise the understanding of molecular mechanism of HCC and also can reveal the protein hub involved in every stage of HCC. Materials and Methods Herein, diethylnitrosamine and thioacetamide (TAA) were used to develop stage-specific HCC in Wistar rats. Histopathological changes, biochemical parameters, and the oxidative stress were measured in hepatocytes. We have reanalyzed the microarray dataset to identify the complex signaling pathways involved in hepatocarcinogenesis induced by TAA. GSE45050 dataset was downloaded from Gene Expression Omnibus database, and the gene expression profile of nontumor, cirrhosis, and HCC was compared. Results The study reveals stage-specific development of chronic HCC rat model and promising stage-specific targets (EHMT2, GMPS, and SPRY2) of HCC. Conclusions EHMT2, GMPS, and SPRY found as high centrality nodes in protein-protein interaction studies using high-throughput microarray data which tend to be present in signaling pathways and co-occur in a biological state of HCC. These genes can be targeted to understand the possible pathology, molecular changes, and target strategy under cirrhosis and HCC condition.
Collapse
Affiliation(s)
- Sandhya Hora
- Department of Biotechnology, HIMT Group of Institutions, Greater Noida; Proteomic and Translational Research Laboratory, Centre for Medical Biotechnology, Amity Institute of Biotechnology, Amity University Uttar Pradesh, Noida, Uttar Pradesh, New Delhi, India
| | - Mohammad Asad
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, Uttar Pradesh, New Delhi, India
| | - Swatantra Kumar Jain
- Department of Biotechnology, School of Chemical and Life Sciences; Hamdard Institute of Medical Sciences and Research, Jamia Hamdard, Uttar Pradesh, New Delhi, India
| | - Deepshikha Pande Katare
- Proteomic and Translational Research Laboratory, Centre for Medical Biotechnology, Amity Institute of Biotechnology, Amity University Uttar Pradesh, Noida, Uttar Pradesh, New Delhi, India
| |
Collapse
|
|
5
|
Ranjpour M, Wajid S, Jain SK. Elevated expression of sepiapterin reductase, regulator of G protein signaling 1, hypothetical protein CXorf58 homolog, and zinc finger and BTB domain-containing protein 21 isoform X2 is associated with progression of hepatocellular carcinoma. PROTOPLASMA 2021; 258:1133-1143. [PMID: 33683453 DOI: 10.1007/s00709-021-01632-2] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/04/2020] [Accepted: 03/01/2021] [Indexed: 06/12/2023]
Abstract
Hepatocellular carcinoma (HCC) is one of the most common cancers associated with high mortality rate. Understanding of events leading to HCC pathophysiology is essential for its better management. We earlier reported development of a novel rodent model by administrating chemical carcinogens, DEN, and 2-AAF for study of HCC at very early stage. 2D-Electrophoresis analysis of total serum proteins identified several differentially expressed proteins in animals undergoing tumorigenesis. MALDI-TOF-MS/MS analyses were performed to characterize the differentially expressed proteins. Further real-time PCR analyses were taken place to quantify the transcript expression for the identified proteins at HCC initiation and tumor stages. Considering protein-protein interactions among the experimentally identified proteins and their interacting neighbors, a protein network has been analyzed that provided further insight into molecular events taking place during HCC development. Histological changes confirmed HCC initiation and hepatotumorigenesis at 1 and 4 months post carcinogen treatment, respectively. Four differentially expressed proteins were identified which were further characterized as regulator of G protein signaling 1 (RGS1), sepiapterin reductase (SPR), similar to zinc finger and BTB domain-containing protein 21 isoform X2 (ZNF295), and a hypothetical protein CXorf58 homolog. Quantification of transcripts for these proteins revealed elevation in their expression both at initiation and tumorigenesis stages. The study deciphers the regulatory role of these proteins during HCC progression.
Collapse
Affiliation(s)
- Maryam Ranjpour
- Departmentof Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, India
| | - Saima Wajid
- Departmentof Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, 110062, India
| | - Swatantra Kumar Jain
- Department of Medical Biochemistry, HIMSR, Jamia Hamdard, New Delhi, 110062, India.
| |
Collapse
|
|
6
|
Castro‐Gil MP, Sánchez‐Rodríguez R, Torres‐Mena JE, López‐Torres CD, Quintanar‐Jurado V, Gabiño‐López NB, Villa‐Treviño S, del‐Pozo‐Jauner L, Arellanes‐Robledo J, Pérez‐Carreón JI. Enrichment of progenitor cells by 2-acetylaminofluorene accelerates liver carcinogenesis induced by diethylnitrosamine in vivo. Mol Carcinog 2021; 60:377-390. [PMID: 33765333 PMCID: PMC8251613 DOI: 10.1002/mc.23298] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/13/2020] [Revised: 02/24/2021] [Accepted: 03/15/2021] [Indexed: 02/06/2023]
Abstract
The potential role of hepatocytes versus hepatic progenitor cells (HPC) on the onset and pathogenesis of hepatocellular carcinoma (HCC) has not been fully clarified. Because the administration of 2-acetylaminofluorene (2AAF) followed by a partial hepatectomy, selectively induces the HPC proliferation, we investigated the effects of chronic 2AAF administration on the HCC development caused by the chronic administration of the carcinogen diethylnitrosamine (DEN) for 16 weeks in the rat. DEN + 2AAF protocol impeded weight gain of animals but promoted prominent hepatomegaly and exacerbated liver alterations compared to DEN protocol alone. The tumor areas detected by γ-glutamyl transferase, prostaglandin reductase-1, and glutathione S-transferase Pi-1 liver cancer markers increased up to 80% as early as 12 weeks of treatment, meaning 6 weeks earlier than DEN alone. This protocol also increased the number of Ki67-positive cells and those of CD90 and CK19, two well-known progenitor cell markers. Interestingly, microarray analysis revealed that DEN + 2AAF protocol differentially modified the global gene expression signature and induced the differential expression of 30 genes identified as HPC markers as early as 6 weeks of treatment. In conclusion, 2AAF induces the early appearance of HPC markers and as a result, accelerates the hepatocarcinogenesis induced by DEN in the rat. Thus, since 2AAF simultaneously administrated with DEN enriches HPC during hepatocarcinogenesis, we propose that DEN + 2AAF protocol might be a useful tool to investigate the cellular origin of HCC with progenitor features.
Collapse
Affiliation(s)
| | - Ricardo Sánchez‐Rodríguez
- Foundation Istituto di Ricerca Pediatrica‐Città della SperanzaPadovaItaly
- Department of Biomedical SciencesUniversity of PadovaPadovaItaly
| | | | | | | | | | - Saúl Villa‐Treviño
- Department of Cell BiologyCenter for Research and Advanced Studies of the National Polytechnic InstituteCiudad de MéxicoMexico
| | | | - Jaime Arellanes‐Robledo
- Laboratory of Liver DiseasesNational Institute of Genomic MedicineCiudad de MéxicoMexico
- Directorate of CátedrasNational Council of Science and TechnologyCiudad de MéxicoMexico
| | | |
Collapse
|
|
7
|
Hasanin AH, Habib EK, El Gayar N, Matboli M. Promotive action of 2-acetylaminofluorene on hepatic precancerous lesions initiated by diethylnitrosamine in rats: Molecular study. World J Hepatol 2021; 13:328-342. [PMID: 33815676 PMCID: PMC8006078 DOI: 10.4254/wjh.v13.i3.328] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/23/2020] [Revised: 12/14/2020] [Accepted: 03/02/2021] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Diethylnitrosamine (DEN) induces hepatic neoplastic lesions over a prolonged period. AIM To investigate the promotive action of 2-acetylaminofluorene (2-AAF) when combined with DEN in order to develop a rat model for induction of precancerous lesion and investigate the molecular mechanism underlying the activity of 2-AAF. METHODS The pre-precancerous lesions were initiated by intraperitoneal injection of DEN for three weeks consecutively, followed by one intraperitoneal injection of 2-AAF at three different doses (100, 200 and 300 mg/kg). Rats were separated into naïve, DEN, DEN + 100 mg 2-AAF, DEN + 200 mg 2-AAF, and DEN + 300 mg 2-AAF groups. Rats were sacrificed after 10 wk and 16 wk. Liver functions, level of alpha-fetoprotein, glutathione S-transferase-P and proliferating cell nuclear antigen staining of liver tissues were performed. The mRNA level of RAB11A, BAX, p53, and Cyclin E and epigenetic regulation by long-noncoding RNA (lncRNA) RP11-513I15.6, miR-1262 (microRNA), and miR-1298 were assessed in the sera and liver tissues of the rats. RESULTS 2-AAF administration significantly increased the percent area of the precancerous foci and cell proliferation along with a significant decrease in RAB11A, BAX, and p53 mRNA, and the increase in Cyclin E mRNA was associated with a marked decrease in lncRNA RP11-513I15.6 expression with a significant increase in both miR-1262 and miR-1298. CONCLUSION 2-AFF promoted hepatic precancerous lesions initiated through DEN by decreasing autophagy, apoptosis, and tumor suppression genes, along with increased cell proliferation, in a time- and dose-dependent manner. These actions were mediated under the epigenetic regulation of lncRNA RP11-513I15.6/miR-1262/miR-1298.
Collapse
Affiliation(s)
- Amany Helmy Hasanin
- Clinical Pharmacology Department, Faculty of Medicine, Ain Shams University, Cairo 11318, Egypt
| | - Eman K Habib
- Anatomy and Embryology Department, Faculty of Medicine, Ain Shams University, Cairo 11318, Egypt
| | - Nesreen El Gayar
- Clinical Pharmacology Department, Faculty of Medicine, Ain Shams University, Cairo 11318, Egypt
| | - Marwa Matboli
- Medical Biochemistry and Molecular Biology Department, Faculty of Medicine, Ain Shams University, Cairo 11381, Egypt.
| |
Collapse
|
|
8
|
Faraj Shaglouf LH, Ranjpour M, Wajid S, Jain SK. Elevated expression of cellular SYNE1, MMP10, and GTPase1 and their regulatory role in hepatocellular carcinoma progression. PROTOPLASMA 2020; 257:157-167. [PMID: 31428857 DOI: 10.1007/s00709-019-01423-w] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/12/2019] [Accepted: 07/19/2019] [Indexed: 06/10/2023]
Abstract
Hepatocellular carcinoma (HCC) is the most common primary liver malignancy resulting in high mortality. HCC progression is associated with abnormal signal transduction that changes cell signaling pathways and ultimately leads to dysregulation of cell functions and uncontrolled cell proliferation. Present study was undertaken with the objective to identify differentially expressed proteins and quantify their transcript expression in the liver of HCC-bearing rats vis-à-vis controls and to decipher the network involving interaction of genes coding for the characterized proteins to an insight into mechanism of HCC tumorigenesis. 2D-Electrophoresis and MALDI-TOF-MS/MS were used to characterize differentially expressed proteins in DEN (diethylnitrosamine)-induced HCC tissue using the protocol reported by us earlier. Real-time PCR was performed to quantify the expression of transcripts for the identified proteins. GENEMANIA, an interacting network of genes coding for selected proteins, was deciphered that provided the functional role of these proteins in HCC progression. Upregulation of proteins SYNE1, MMP10, and MTG1 was observed. The mRNA quantification revealed elevated expression of their transcripts at HCC initiation, progression, and tumor stages. Network analysis showed the involvement of the genes coding for these proteins in dysregulation of signaling pathways during HCC development. The elevated expression of SYNE1, MMP10, and MTG1 suggests the role of these proteins as potential players in HCC progression and tumorigenesis.
Collapse
Affiliation(s)
- Laila H Faraj Shaglouf
- Department of Biotechnology, School of Chemical and Life Science, Jamia Hamdard, New Delhi, 110062, India
| | - Maryam Ranjpour
- Department of Biotechnology, School of Chemical and Life Science, Jamia Hamdard, New Delhi, 110062, India
| | - Saima Wajid
- Department of Biotechnology, School of Chemical and Life Science, Jamia Hamdard, New Delhi, 110062, India
| | - Swatantra Kumar Jain
- Department of Biochemistry, Hamdard Institute of Medical Science and Research, Jamia Hamdard, New Delhi, 110062, India.
| |
Collapse
|
|
9
|
Mohamed Y, Basyony MA, El-Desouki NI, Abdo WS, El-Magd MA. The potential therapeutic effect for melatonin and mesenchymal stem cells on hepatocellular carcinoma. Biomedicine (Taipei) 2019; 9:24. [PMID: 31724939 PMCID: PMC6855194 DOI: 10.1051/bmdcn/2019090424] [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: 05/09/2019] [Accepted: 06/25/2019] [Indexed: 12/11/2022] Open
Abstract
BACKGROUND/AIM Herein, we investigated the potential therapeutic effect of Melatonin (Mel) and/or mesenchymal stem cells (MSCs) on rat model of HCC. MATERIALS AND METHODS Female mature rats were divided into 5 groups (n = 10/group): normal (Nor), HCC group intraperitoneally injected with 200 mg/kg DEN, and 3 treated groups; HCC + Mel (Mel) group given Mel intraperitoneally 20 mg/kg, twice a week, HCC + MSCs (MSCs) group intravenously injected by 1 × 106 cells, and HCC + MSCs (Mel +MSCs) group. RESULTS Rats in HCC group showed most deteriorated effect in form of increased mortality and relative liver weight, elevated serum levels of ALT, AST, ALP, AFP and GGT in addition to increased pre-neoplastic nodules in liver tissues. Liver tissues of HCC group also exhibited lower level of apoptosis as indicated by decreased DNA fragmentation and expression of p53 caspase 9 and caspase 3 genes and increased PCNA immunoreactivity. Moreover, in this group the expression of IL6 and TGFβ1 genes was significantly upregulated. All these deleterious effects induced by DEN were reversed after administration of Mel and/ or MSCs with best improvement for the combined group (MSCs + Mel). CONCLUSIONS These findings reveal a better therapeutic effect for MSCs when given with Mel and we attribute this beneficial effect, at least in part, to triggering apoptosis and targeting inflammation in HCC. Therefore, combined treatment with Mel and MSCs is recommended to enhance the therapeutic potential against HCC.
Collapse
Affiliation(s)
- Yasser Mohamed
- Department of Zoology, Faculty of Science, Tanta University, Tanta 31527, Egypt
| | - Mohamed A Basyony
- Department of Zoology, Faculty of Science, Tanta University, Tanta 31527, Egypt
| | - Nabila I El-Desouki
- Department of Zoology, Faculty of Science, Tanta University, Tanta 31527, Egypt
| | - Walied S Abdo
- Department of Pathology, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh 33516, Egypt
| | - Mohammed A El-Magd
- Department of Anatomy, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh 33516, Egypt
| |
Collapse
|
|
10
|
Ranjpour M, Wajid S, Jain SK. Elevated Expression of A-Raf and FA2H in Hepatocellular Carcinoma is Associated with Lipid Metabolism Dysregulation and Cancer Progression. Anticancer Agents Med Chem 2019; 19:236-247. [PMID: 30324893 DOI: 10.2174/1871520618666181015142810] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2017] [Revised: 05/08/2018] [Accepted: 09/25/2018] [Indexed: 02/07/2023]
Abstract
BACKGROUND Identification of events leading to hepatocellular carcinoma (HCC) progression is essential for understanding its pathophysiology. The aims of this study are to identify and characterize differentially expressed proteins in serum of HCC-bearing rats and the corresponding controls during cancer initiation, progression and tumorigenesis. METHODS Chemical carcinogens, N-Nitrosodiethylamine and 2-aminoacetylfluorine are administered to induce HCC to male Wistar rats. The 2D-Electrophoresis and PD-Quest analyses are performed to identify several differentially expressed proteins in serum of HCC-bearing animals. These proteins are further characterized by MALDI-TOF-MS/MS analyses. Using pathwaylinker a HCC-specific network is analyzed among the MALDITOF- MS/MS characterized proteins and their interactors. RESULTS Carcinogen administration caused inflammation leading to liver injury and HCC development. Liver inflammation was confirmed by increase in the levels of TNF-α and IL-6 in carcinogen treated rats. We report significant increase in expression of two differentially expressed proteins, namely, A-Raf and Fatty Acid 2- Hydroxylase (FA2H), at early stage of HCC initiation, during its progression and at tumor stage. Real-time PCR analysis of mRNA for these proteins confirmed up-regulation of their transcripts. Further, we validated our experimental data with sera of clinically confirmed liver cancer patients. CONCLUSION The study suggests that FA2H and A-Raf play a major role in the progression of HCC.
Collapse
Affiliation(s)
- Maryam Ranjpour
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi 110062, India
| | - Saima Wajid
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi 110062, India
| | - Swatantra K Jain
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi 110062, India.,Department of Medical Biochemistry, HIMSR, Jamia Hamdard, New Delhi 110062, India
| |
Collapse
|
|
11
|
Ranjpour M, Wajid S, Jain SK. Elevated Expression of Cytosolic Phospholipase A2 Delta Is Associated with Lipid Metabolism Dysregulation during Hepatocellular Carcinoma Progression. CELL JOURNAL 2019; 22:17-22. [PMID: 31606962 PMCID: PMC6791066 DOI: 10.22074/cellj.2020.6527] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/06/2018] [Accepted: 02/04/2019] [Indexed: 12/15/2022]
Abstract
Objective Liver cancer is the third rank amongst the common malignancies, causing maximum death in the patients
diagnosed with cancers. Currently available biomarkers are not enough sensitive for early diagnosis of hepatocellular
carcinoma (HCC). This makes difficult management of HCC. With the aim of finding new generation of proteomic-based
biomarkers, the represented study was designed to characterize the differentially expressed proteins at different stages
of HCC initiation and at progression. This could lead to find potential biomarkers for early detection of HCC.
Materials and Methods In this experimental study, we report induction of HCC by administrating chemical carcinogens
in male Wistar rats. Disease progression was monitored by histological evaluation. Serum proteomic analyses such as
2 dimensional (2D)-electrophoresis, MALDI-TOF-MS/MS and Western blot have been used to analyze and characterize
the differentially expressed proteins during HCC development.
Results HCC initiation and tumorigenesis were observed at one and four months post carcinogen treatment,
respectively. One of the differentially-expressed proteins, namely, cytosolic phospholipase A2delta was significantly
up-regulated at very early stage of HCC development. Its expression continued to increase during cancer progression
and hepatotumorigenesis stages. Its elevated expression has been confirmed by Western blot analysis. Consistent to
this, analyses of the sera in the clinically confirmed liver cancer patients showed elevated expression of this protein,
further validating our experimental results.
Conclusion This study suggests that elevation in the expression of cytosolic phospholipase A2delta is associated with
progression of HCC.
Collapse
Affiliation(s)
- Maryam Ranjpour
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, India
| | - Saima Wajid
- Department of Biotechnology, School of Chemical and Life Sciences, Jamia Hamdard, New Delhi, India
| | - Swatantra Kumar Jain
- Department of Medical Biochemistry, HIMSR, Jamia Hamdard, New Delhi, India. Electronic Address:
| |
Collapse
|
|
12
|
El-Magd MA, Mohamed Y, El-Shetry ES, Elsayed SA, Abo Gazia M, Abdel-Aleem GA, Shafik NM, Abdo WS, El-Desouki NI, Basyony MA. Melatonin maximizes the therapeutic potential of non-preconditioned MSCs in a DEN-induced rat model of HCC. Biomed Pharmacother 2019; 114:108732. [PMID: 30925457 DOI: 10.1016/j.biopha.2019.108732] [Citation(s) in RCA: 36] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2018] [Revised: 02/22/2019] [Accepted: 02/22/2019] [Indexed: 12/21/2022] Open
Abstract
Pretreatment of mesenchymal stem cells (MSCs) with melatonin (Mel) improves their potential therapeutic effect on chronic diseases and cancers. However, this preconditioning strategy may direct the effect of Mel toward MSCs alone and deprive cancer cells of the oncostatic effect of Mel. Herein, we hypothesized that Mel given before transplantation of non-preconditioned MSCs may maximize the therapeutic outcome via the oncostatic effect of Mel by preparing a suitable tumor microenvironment for MSCs. Female rats (n = 60) were equally divided into 6 groups; normal control, diethylnitrosamine (DEN), DEN + Mel, DEN + MSCs, DEN + MSCs preconditioned with Mel, and DEN + MSCs + Mel. The obtained data revealed that administration of Mel before MSCs treatment without preconditioning yielded a better ameliorative effect against DEN-induced hepatocellular carcinoma (HCC) as evidenced by: 1) reduced serum levels of alpha fetoprotein and gamma-glutamyl transferase; 2) decreased number and area of glutathione S-transferase placental positive foci; 3) induced apoptosis (as indicated by increased cleaved caspase-3 activity, upregulated expression of proapoptotic genes Bax and caspase 3 and downregulated expression of anti-apoptotic genes Bcl2, survivin); 4) decreased malondialdehyde level and increased activities of superoxide dismutase, catalase, and glutathione peroxidase enzymes; and 5) reduced inflammation, angiogenesis and metastasis as indicated by downregulated expression of interleukin 1 beta, nuclear factor kappa B, vascular endothelial growth factor, and matrix metallopeptidase 9 genes and upregulated expression of metalloproteinase inhibitor 1 gene. Thus, administration of Mel before MSCs (without preconditioning) fostered the survival and therapeutic potential of MSCs in HCC, possibly through induction of apoptosis and inhibition of inflammation and oxidative stress. This new strategy showed better therapeutic outcomes and may improve MSC-based therapies for HCC.
Collapse
Affiliation(s)
- Mohammed A El-Magd
- Department of Anatomy, Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt.
| | - Yasser Mohamed
- Department of Zoology, Faculty of Science, Tanta University, Egypt
| | - Eman S El-Shetry
- Department of Anatomy, Faculty of Medicine, Zagazig University, Egypt
| | - Shafika A Elsayed
- Department of Histology and Cytology, Faculty of Veterinary Medicine, Zagazig University, Egypt
| | - Maha Abo Gazia
- Department of Histology, Faculty of Medicine, Kafrelsheikh University, Egypt
| | - Ghada A Abdel-Aleem
- Department of Medical Biochemistry, Faculty of Medicine, Tanta University, Egypt
| | - Noha M Shafik
- Department of Medical Biochemistry, Faculty of Medicine, Tanta University, Egypt
| | - Walied S Abdo
- Department of Pathology, Faculty of Veterinary Medicine, Kafrelsheikh University, Egypt
| | | | | |
Collapse
|
|
13
|
Thomas NS, George K, Selvam AAA. Troxerutin subdues hepatic tumorigenesis via disrupting the MDM2-p53 interaction. Food Funct 2019; 9:5336-5349. [PMID: 30259932 DOI: 10.1039/c8fo01111g] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
Abstract
Hepatocellular carcinoma (HCC) is the leading cause of cancer death worldwide that lacks proper medical prognosis and treatment. In the present study, the anti-tumoral potential of troxerutin (TX), an ethnomedicine, was examined in relation to its effects on the promoter 2-acetylaminofluorene (2-AAF) in N-nitrosodiethylamine (NDEA) initiated HCC, as compared to its effects on HCC induced by NDEA alone. Liver samples from each experimental group were collected and evaluated for histological, biochemical and cellular characterization. The protein expressions of apoptotic and cell proliferation markers were determined via immunohistochemistry and western blotting. Molecular docking was also performed to delineate the inhibitory mechanism of TX on HCC. The results show that only higher doses of TX showed a significant reduction in the incidence of hepatic nodule formation, and they also counteracted NDEA plus 2-AAF induced alterations in the enzymic status. The frequencies of glutathione-S-transferase and proliferating cell nuclear antigen, markers of S phase progression, were markedly reduced during TX treatment. TX also modulated the imbalance in the MDM2-p53 interaction. The molecular docking results confirmed the interaction of TX with the upstream kinases that regulate apoptosis. This study provides evidence that a copious dose of TX is required to counteract the differential mitoinhibitory effect of 2-AAF in NDEA initiated hepatomas, and TX exhibits an anti-tumoral effect via suppressing oxidative stress, regulating liver function enzymes, inhibiting inflammatory responses and modulating MDM2-p53 interactions, thus inducing apoptosis, and thereby suggesting that TX may provide promising therapeutic effects for the chemoprevention of HCC.
Collapse
Affiliation(s)
- Nisha Susan Thomas
- Department of Biochemistry and Biotechnology, Annamalai University, Tamil Nadu, India.
| | | | | |
Collapse
|
|
14
|
Effect of diosmin on apoptotic signaling molecules in N-nitrosodiethylamine-induced hepatocellular carcinoma in experimental rats. Mol Cell Biochem 2018; 449:27-37. [PMID: 29479636 DOI: 10.1007/s11010-018-3339-3] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2017] [Accepted: 02/22/2018] [Indexed: 12/22/2022]
Abstract
The aim of the present study was to evaluate the antioxidant and chemopreventive efficiency of diosmin against N-nitrosodiethylamine (NDEA)-induced hepatocarcinogenesis in adult male rats. Rats were classified into four groups as follows: Group I: Control, Group II: NDEA-induced hepatocellular carcinogenic rats, Group III: Cancer-bearing animals treated with diosmin (200 mg/kg/body weight/day) orally for 28 days, Group IV: Control animals treated with diosmin (200 mg/kg/body weight/day) alone for 28 days. The model of NDEA-induced HCC rats elicited significant increases in alpha-fetoprotein (AFP), lipid peroxidation (LPO) and increase in anti-apoptotic proteins (Bcl-2, Bcl-xL and Mcl-1) with a concomitant significant decline in liver antioxidant enzymes, pro-apoptotic (Bax and Bad) and caspase-3 &-9 proteins. The oral administration of diosmin as a protective agent normalized the altered levels of AFP, LPO, antioxidant enzymes, pro- and anti-apoptotic proteins as well as caspase-3 and -9 proteins. Transmission electron microscopical studies also revealed that treatment of diosmin has a perspective anti-cancer activity by rearranging hepatic cell structure and its integrity. Results of this study suggest that diosmin may be one of a pharmacological and therapeutic representative against hepatocellular carcinoma.
Collapse
|
|
15
|
P. Katare D, Malik S, J. Mani R, Ranjpour M, Jain SK. Novel mutations in transthyretin gene associated with hepatocellular carcinoma. Mol Carcinog 2017; 57:70-77. [DOI: 10.1002/mc.22732] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/25/2017] [Revised: 08/08/2017] [Accepted: 09/05/2017] [Indexed: 02/04/2023]
Affiliation(s)
- Deepshikha P. Katare
- Proteomics and Translational Research Lab; Centre for Medical Biotechnology; Amity Institute of Biotechnology; Amity University; Noida Uttar Pradesh India
| | - Shabnam Malik
- Faculty of Chemical and Life Sciences; Department of Biotechnology; Hamdard Institute of Medical Sciences and Research; Hamdard University; New Delhi India
| | - Ruchi J. Mani
- Proteomics and Translational Research Lab; Centre for Medical Biotechnology; Amity Institute of Biotechnology; Amity University; Noida Uttar Pradesh India
| | - Maryam Ranjpour
- Faculty of Chemical and Life Sciences; Department of Biotechnology; Hamdard Institute of Medical Sciences and Research; Hamdard University; New Delhi India
| | - Swatantra K. Jain
- Faculty of Chemical and Life Sciences; Department of Biotechnology; Hamdard Institute of Medical Sciences and Research; Hamdard University; New Delhi India
- Department of Medical Biochemistry; Hamdard Institute of Medical Sciences and Research; Hamdard University; New Delhi India
| |
Collapse
|
|
16
|
Moreira AJ, Rodrigues GR, Bona S, Fratta LXS, Weber GR, Picada JN, Dos Santos JL, Cerski CT, Marroni CA, Marroni NP. Ductular reaction, cytokeratin 7 positivity, and gamma-glutamyl transferase in multistage hepatocarcinogenesis in rats. PROTOPLASMA 2017; 254:911-920. [PMID: 27525410 DOI: 10.1007/s00709-016-1000-0] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/31/2015] [Accepted: 06/22/2016] [Indexed: 06/06/2023]
Abstract
Hepatocellular carcinoma (HCC) is the most common primary malignancy of the liver and is characterized by multistage formation. The presence of ductular reaction, cytokeratin 7 positivity (PCK7), and increased levels of gamma glutamyltransferase (γGT) has been observed during liver carcinogenesis and contribute to tumor progression. Our goal was to evaluate the ductular reaction in multistage carcinogenesis and to correlate PCK7 and γGT levels with tumor incidence, histological characteristics, liver DNA damage index, and the expression of oxidative stress proteins. HCC was induced in 24 male Wistar rats weighing 145-150 g by chronic and intermittent exposure to 50 or 100 mg/kg diethylnitrosamine (DEN). Six control animals received only vehicle. Blood was collected to determine hepatic enzyme levels. Animals were divided into three groups: control (CO), precancerous lesions (PL), and advanced HCC. Liver samples were obtained for immunohistochemical analyses and the measurement of protein expression. Statistical analyses included Tukey's test and Pearson's correlation analyses. We observed an extensive ductular reaction in advanced HCC and a strong correlation between PCK7 and levels of γGT and the poor prognosis and aggressiveness of HCC. The extent of PCK7 and high γGT levels were associated with overexpression of inducible nitric oxide synthase (iNOS) and heat shock factor protein 1 (HSF-1). However, PCK7 and γGT levels were negatively correlated with protein expression of nuclear factor erythroid 2-related factor 2 (NRF2) and inducible heat shock protein 70 (iHSP70). These findings suggest that ductular reaction is involved in the progression of multistage hepatocarcinogenesis.
Collapse
Affiliation(s)
- Andrea Janz Moreira
- Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil
- Department of Biological Sciences: Physiology, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500, Porto Alegre, RS, Brazil
- Department of Physical Therapy of Porto Alegre Institute, IPA, Rua Joaquim Pedro Salgado, 80, Porto Alegre, Brazil
| | - Graziella Ramos Rodrigues
- Gene Therapy Center, Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, Brazil
| | - Silvia Bona
- Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil
| | - Leila Xavier Sinigaglia Fratta
- Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil
| | - Giovana Regina Weber
- Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil
| | - Jaqueline Nascimento Picada
- Program in Cell and Molecular Biology Applied to Health, Universidade Luterana do Brasil, Av. Farroupilha, 8001, Canoas, Brazil
| | - Jorge Luiz Dos Santos
- Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil
- Pediatric Hepatology Unit, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, Brazil
| | - Carlos Thadeu Cerski
- Department of Pathology, School of Medicine, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
| | - Claudio Augusto Marroni
- Program in Liver Diseases, Universidade Federal de Ciências da Saúde de Porto Alegre, Rua Sarmento Leite, 245, Porto Alegre, Brazil
| | - Norma Possa Marroni
- Center of Experimental Research, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil.
- Department of Biological Sciences: Physiology, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite, 500, Porto Alegre, RS, Brazil.
- Program in Cell and Molecular Biology Applied to Health, Universidade Luterana do Brasil, Av. Farroupilha, 8001, Canoas, Brazil.
- , Rua José Kanan Aranha 102, 91760-470, Porto Alegre, RS, Brazil.
| |
Collapse
|
|
17
|
The in vivo antineoplastic and therapeutic efficacy of troxerutin on rat preneoplastic liver: biochemical, histological and cellular aspects. Eur J Nutr 2016; 56:2353-2366. [PMID: 27488610 DOI: 10.1007/s00394-016-1275-0] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2016] [Accepted: 07/11/2016] [Indexed: 12/18/2022]
Abstract
PURPOSE Troxerutin (TXER), a trihydroxyethylated derivative of the natural bioflavonoid rutin, abundantly found in tea, various fruits and vegetables, is known to exhibit ample pharmacological properties. In the present investigation, we examined the antineoplastic, therapeutic efficacy and furthermore the possible mechanisms of action of TXER against NAFLD/NASH progression to hepatocarcinogenesis. METHODS The effect of TXER (12.5, 25 or 50 mg/kg b.w/day) was evaluated on the nitrosodiethylamine (NDEA) model of hepatocarcinogenesis in rats, after 16 weeks of oral treatment, with special focus on liver specific enzymes, xenobiotic metabolizing enzymes, antioxidant status, lipid peroxidation profile, DNA damage, fibrosis, cell proliferation and inflammatory status. RESULTS Administration of TXER to hepatocellular carcinoma-bearing rats restored the enzyme activities and the hepatic architecture. Furthermore, TXER significantly curtailed NDEA-induced DNA damage, cell proliferation, inflammation, fibrosis and hepatic hyperplasia. CONCLUSION This study provides the evidence that troxerutin exerts a significant therapeutic effect against liver cancer by modulating liver function enzymes, xenobiotic enzymes, oxidative damage, inhibiting cell proliferation, suppressing inflammatory response and induction of apoptosis.
Collapse
|
|
18
|
Bona S, Moreira AJ, Rodrigues GR, Cerski CT, da Silveira TR, Marroni CA, Marroni NP. Diethylnitrosamine-induced cirrhosis in Wistar rats: an experimental feasibility study. PROTOPLASMA 2015; 252:825-833. [PMID: 25369754 DOI: 10.1007/s00709-014-0719-8] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/19/2014] [Accepted: 10/13/2014] [Indexed: 06/04/2023]
Abstract
The experimental models of the development of cirrhosis in rats require a long time. Many studies in animals have demonstrated similarities in histological pattern with human cirrhosis. Just like the relation between cirrhosis and increased lipid peroxidation (LPO), which contributes to the worsening of the disease. However, few studies have focused on the reduction of time to establish cirrhosis and evaluated the expression of heat-shock protein 70 (HSP70) in cirrhotic livers of rodents. The present study proposes the adaptation of an experimental cirrhosis model using diethylnitrosamine (DEN). Twenty-six male Wistar rats, weighing ±270 g, divided into two groups: (i) CO-control and (ii) DEN-diethylnitrosamine. The DEN group received 50 mg/kg of DEN twice a week intraperitoneally for 7 weeks. The model developed cirrhosis in 7 weeks. The liver function tests showed that the animals with DEN-induced cirrhosis had increased levels when compared to control. The histological examination showed changes in the liver architecture, with severe ductal proliferation, signs of chronic damage, cholestasis, lymphocytic infiltrate, steatosis, and extensive parenchymal loss. We also found nodular formations with homogeneous pattern, increased LPO, increased expression of iNOS, TGF beta, α-SMA, and NQO1. However, the HSP70 expression was reduced in cirrhotic animals. This study showed signs of cirrhosis in liver based on biochemical, histological, and molecular analysis. The reduced expression of HSP70 appears to be associated with increased oxidative stress, contributing to the worsening of the disease.
Collapse
Affiliation(s)
- Silvia Bona
- Postgraduate Program in Medical Sciences: Medicine, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2400, Porto Alegre, RS, Brazil
| | | | | | | | | | | | | |
Collapse
|
|
19
|
Oxidative stress and cell damage in a model of precancerous lesions and advanced hepatocellular carcinoma in rats. Toxicol Rep 2014; 2:333-340. [PMID: 28962366 PMCID: PMC5598147 DOI: 10.1016/j.toxrep.2014.11.015] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2014] [Revised: 11/21/2014] [Accepted: 11/21/2014] [Indexed: 12/20/2022] Open
Abstract
Hepatocellular carcinoma (HCC) is the third most frequent cause of cancer deaths throughout the world. This study was aimed to analyze oxidative stress and cell damage in a multistage model of liver carcinogenesis induced by diethylnitrosamine (DEN) in rats. Male Wistar rats weighing 145–150 g were divided into three groups: control, precancerous lesions (PL) (which received 100 mg DEN once a week every 6 weeks up to 28 weeks), and advanced HCC (50 mg DEN once/twice per week up to 19 weeks). Lipid peroxidation (TBARS), superoxide dismutase (SOD) activity, and expression of transforming growth factor-1 beta (TGF)-1β, endothelial and inducible nitric oxide syntahese (eNOS, iNOS), NADPH quinone oxireductase (NQO)-1, nuclear factor erythroid 2-related factor (NrF)2, kelch-like ECH-associated protein (Keap)1 and heat shock protein (HSP)70 were measured. TBARS concentration was augmented in the PL and advanced HCC groups. SOD activity, TGF-1β and Nrf2 expression were higher in animals with precancerous lesions. In advanced HCC, expression of NQO1 and iNOS increased while there was a decrease in HPS70 expression. Data obtained provide evidence for the differential activation of proteins involved in oxidative stress and cell damage during progression of carcinogenesis in an animal model of HCC.
Collapse
Key Words
- 2-AAF, 2-acetylaminofluorene
- ALT, alanine aminotransferase
- AP, alkaline phosphatase
- AST, aspartate aminotransferase
- DEN, diethylnitrosamine
- Diethylnitrosamine
- EDTA, ethylenediamine tetraacetic acid
- GGT, gamma-glutamyl transferase
- HCC, hepatocellular carcinoma
- HSC, hepatic stellate cells
- HSP70, heat shock 70-kDa protein
- Heat shock protein
- Hepatocarcinoma
- Keap1, kelch-like ECH-associated protein 1
- MDA, malonaldehyde
- NO, nitric oxide
- NQO1, NADPH quinone oxireductase-1
- Nitric oxide synthase
- Nrf2, nuclear factor erythroid 2-related factor 2
- Nuclear factor erythroid 2-related factor 2
- Oxidative stress
- PVDF, polyvinylidene fluoride
- SOD, superoxide dismutase
- TBARS, thiobarbituric acid reactant substances
- TGF-1β, transforming growth fator-1 beta
- TTBS, Tris-buffered containing 0.05% Tween 20
- UV, ultra violet
- eNOS, endothelial nitric oxide synthase
- iNOS, inducible nitric oxide synthase
Collapse
|
|
20
|
Alam S, Yadav RS, Pal A, Purshottam SK, Chaudhari BP, Das M, Ansari KM. Dietary administration of Nexrutine inhibits rat liver tumorigenesis and induces apoptotic cell death in human hepatocellular carcinoma cells. Toxicol Rep 2014; 2:1-11. [PMID: 28962332 PMCID: PMC5598519 DOI: 10.1016/j.toxrep.2014.11.006] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/13/2014] [Revised: 11/04/2014] [Accepted: 11/04/2014] [Indexed: 01/14/2023] Open
Abstract
Nexrutine has anti-tumor potential in Solt-Farber rat liver tumorigenesis model. Nexrutine caused decreased cell proliferation in the DEN/2-AAF treated rats. It decreases cell viability of liver cancer cells and modulates pro- and anti-apoptotic markers. Nexrutine modulates the cell cycle regulatory proteins and MAPKs. Epidemiological studies suggested that plant-based dietary supplements can reduce the risk of liver cancer. Nexrutine (NX), an herbal extract from Phellodendronamurense, has been shown to have anti-inflammatory, anti-microbial and anti-tumor activities. In the present study, we have shown the anti-tumor potential of NX against Solt-Farber model with elimination of PH, rat liver tumor induced by diethylnitrosoamine (DEN) as carcinogen and 2-acetylaminofluorene (2-AAF) as co-carcinogen. The elucidation of mechanistic pathways was explored in human liver cancer cells. Dietary intake of NX significantly decreased the cell proliferation and inflammation, as well as increased apoptosis in the liver sections of DEN/2-AAF-treated rats. Moreover, NX (2.5–10 μg/ml) exposure significantly decreased the viability of liver cancer cells and modulated the levels of Bax and Bcl-2 proteins levels. NX treatment resulted in increased cytochrome-c release and cleavage of caspases 3 and 9. In addition, NX decreased the expression of CDK2, CDK4 and associated cyclins E1 and D1, while up-regulated the expression of p21, p27 and p53 expression. NX also enhanced phosphorylation of the mitogen-activated protein kinases (MAPKs) ERK1/2, p38 and JNK1/2. Collectively, these findings suggested that NX-mediated protection against DEN/2-AAF-induced liver tumorigenesis involves decrease in cell proliferation and enhancement in apoptotic cell death of liver cancer cells.
Collapse
Affiliation(s)
- Shamshad Alam
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| | - Ravi S Yadav
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| | - Anu Pal
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| | - Shakendra K Purshottam
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| | - Bhushan P Chaudhari
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| | - Mukul Das
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| | - Kausar M Ansari
- Food, Drug and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (CSIR-IITR), Mahatma Gandhi Marg, P.O. Box#80, Lucknow 226001, India
| |
Collapse
|
|
21
|
Sikander M, Malik S, Parveen K, Ahmad M, Yadav D, Hafeez ZB, Bansal M. Hepatoprotective effect of Origanum vulgare in Wistar rats against carbon tetrachloride-induced hepatotoxicity. PROTOPLASMA 2013; 250:483-493. [PMID: 22772591 DOI: 10.1007/s00709-012-0431-5] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/04/2012] [Accepted: 06/26/2012] [Indexed: 06/01/2023]
Abstract
The effect of an aqueous extract of Origanum vulgare (OV) leaves extract on CCl4-induced hepatotoxicity was investigated in normal and hepatotoxic rats. To evaluate the hepatoprotective activity of OV, rats were divided into six groups: control group, O. vulgare group, carbon tetrachloride (CCl4; 2 ml/kg body weight) group, and three treatment groups that received CCl4 and OV at doses of 50, 100, 150 mg/kg body weight orally for 15 days. Alanine amino transferase (ALT), alkaline phosphatase (ALP), and aspartate amino transferase (AST) in serum, lipid peroxide (LPO), GST, CAT, SOD, GPx, GR, and GSH in liver tissue were estimated to assess liver function. CCl4 administration led to pathological and biochemical evidence of liver injury as compared to controls. OV administration led to significant protection against CCl4-induced hepatotoxicity in dose-dependent manner, maximum activity was found in CCl4 + OV3 (150 mg/kg body weight) groups and changes in the hepatocytes were confirmed through histopathological analysis of liver tissues. It was also associated with significantly lower serum ALT, ALP, and AST levels, higher GST, CAT, SOD, GPx, GR, and GSH level in liver tissue. The level of LPO also decreases significantly after the administration of OV leaves extract. The biochemical observations were supplemented with histopathological examination of rat liver sections. Thus, the study suggests O. vulgare showed protective activity against CCl4-induced hepatotoxicity in Wistar rats and might be beneficial for the liver toxicity.
Collapse
Affiliation(s)
- Mohammad Sikander
- Department of Biotechnology, Hamdard University, Hamdard Nagar, New Delhi, 110062, India.
| | | | | | | | | | | | | |
Collapse
|
|
22
|
Elevated expression of complement C3 protein in chemically induced hepatotumorogenesis in Wistar rats: a correlative proteomics and histopathological study. ACTA ACUST UNITED AC 2012; 65:767-73. [PMID: 23245919 DOI: 10.1016/j.etp.2012.11.003] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/03/2012] [Accepted: 11/08/2012] [Indexed: 11/20/2022]
Abstract
Liver cancer remains the leading cause of cancer-related mortality worldwide. Early detection of liver cancer is problematic due to the lack of a marker with high diagnosis sensitivity and specificity. The present study was designed to determine the differently expressed proteins at early stage in the serum of animals with liver cancer vis-à-vis controls and figure out the function of the proteins. One-dimensional electrophoresis (1D), two-dimensional electrophoresis (2DE) and liquid chromatography mass spectrometry (LC-MS/MS) were used to screen the serum proteins of liver cancer induced in animals by diethyl nitrosamine (DEN)+2-acetyl amino fluorine (2-AAF). From optimized 2DE image and computer assisted PD Quest analysis were found to be differentially expressed spots when the serum from normal and treated animals were compared. Among these, one spot was selected whose expression level was higher in DEN+2-AAF treated animal sera than in adjacent normal animal sera. The target spot was excised from the 2D gel of liver cancer sera and the peptide mass fingerprinting as obtained LC-MS/MS analysis after digesting the chosen protein spot. This was identified to be complement C3 protein. The changes in complement C3 expression level were validated by Western blot analysis. We reported that the changes in complement C3 concentration start at very early stage of tumorogenesis. The fully grown tumors were developed at 120 days and hepatotumorogenesis was confirmed by histopathological examination. This protein may therefore represent a powerful tool in search for candidate biomarkers for HCC.
Collapse
|