|
1
|
Inhibition of MZF1/c-MYC Axis by Cantharidin Impairs Cell Proliferation in Glioblastoma. Int J Mol Sci 2022; 23:ijms232314727. [PMID: 36499054 PMCID: PMC9740304 DOI: 10.3390/ijms232314727] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/28/2022] [Revised: 11/15/2022] [Accepted: 11/23/2022] [Indexed: 11/27/2022] Open
Abstract
Myeloid zinc finger 1 (MZF1), also known as zinc finger protein 42, is a zinc finger transcription factor, belonging to the Krüppel-like family that has been implicated in several types of malignancies, including glioblastoma multiforme (GBM). MZF1 is reportedly an oncogenic gene that promotes tumor progression. Moreover, higher expression of MZF1 has been associated with a worse overall survival rate among patients with GBM. Thus, MZF1 may be a promising target for therapeutic interventions. Cantharidin (CTD) has been traditionally used in Chinese medicine to induce apoptosis and inhibit cancer cell proliferation; however, the mechanism by which CTD inhibits cell proliferation remains unclear. In this study, we found that the expression of MZF1 was higher in GBM tissues than in adjacent normal tissues and low-grade gliomas. Additionally, the patient-derived GBM cells and GBM cell lines presented higher levels of MZF1 than normal human astrocytes. We demonstrated that CTD had greater anti-proliferative effects on GBM than a derivative of CTD, norcantharidin (NCTD). MZF1 expression was strongly suppressed by CTD treatment. Furthermore, MZF1 enhanced the proliferation of GBM cells and upregulated the expression of c-MYC, whereas these effects were reversed by CTD treatment. The results of our study suggest that CTD may be a promising therapeutic agent for patients with GBM and suggest a promising direction for further investigation.
Collapse
|
|
2
|
Synthesis and bioactivity evaluation of 5,6-epoxynorcantharidin mono-amide and imide derivatives. MONATSHEFTE FUR CHEMIE 2022. [DOI: 10.1007/s00706-022-02905-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
|
|
3
|
Tan A, Kizilkaya S, Kelestemur U, Akdemir A, Kara Y. The Synthesis, Anticancer Activity, Structure-Activity Relationships and Molecular Modelling Studies of Novel Isoindole-1,3(2H)-dione Compounds Containing Different Functional Groups. Anticancer Agents Med Chem 2020; 20:1368-1378. [DOI: 10.2174/1871520620666200410080648] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2019] [Revised: 02/20/2020] [Accepted: 03/02/2020] [Indexed: 11/22/2022]
Abstract
Background:
Isoindole-1,3(2H)-dione derivatives are known to have cytotoxic effects on many
cancer cells. The anticancer activity of these compounds varies depending on the substituents attached to them.
Therefore, the effect of substituents is very important when determining the anticancer activities of molecules.
We have recently reported an example of the substituent effect.
:
According to that work, the anticancer activity against HeLa, C6, and A549 cancer cell lines of isoindole-
1,3(2H)-dione compounds containing tert-butyldiphenylsilyl ether, azido, and hydroxyl groups was examined by
our group. It was found that an isoindole-1,3(2H)-dione compound containing both tert-butyldiphenylsilyl ether
group and azido groups showed higher anticancer activity than 5-fluorouracil and another isoindole-1,3(2H)-
dione compound containing both azido and hydroxyl groups.
:
After we discovered that tert-butyldiphenylsilyl ether group in the skeletal structure of isoindole-1,3(2H)-dione
exhibits anticancer activity against HeLa, C6, and A549 cancer cell lines, we wanted to examine the anticancer
activities of different silyl ether groups, i.e., OTMS, -OTBDPS, and -OTBDMS groups, and also -OH and -Br
groups, by comparing them with each other according to the structure–activity relationship.
Methods:
All of the synthesized compounds were characterized by 1H and 13C NMR spectra, IR spectroscopy,
and mass spectra measurements. The IC50 values of these compounds were calculated for all cancer cell lines
and compared with each other and cisplatin, which is a platinum-containing chemotherapeutic drug. Molecular
modelling studies were carried out using the MOE software package.
Results:
It was found that compounds 13 and 16, containing both silyl ether (-OTBDMS) and -Br groups,
showed higher anticancer activity than cisplatin against both Caco-2 and MCF-7 cell lines. Compounds 20 and
23 showed anticancer activity in MCF-7 cells and compounds 8, 9, 20, and 23 in Caco-2 cells. While
compounds 20 and 23 have only a silyl ether (-OTMS) group, compounds 8 and 9 have only a -OH group.
Molecular modelling studies indicated that compounds 8 and 13, as well as their analogs, may bind to the active
site of hRS6KB1 (pdb: 4l3j), compound 11 may bind to the active site of human mTOR (pdb: 4jt5) and
additionally, compounds 10-17 are expected to be both mutagenic and reactive according to the mutagenicity
and reactivity calculations.
Conclusion:
According to these results, the anticancer activities of isoindole-1,3(2H)-dione compounds (8 - 23)
vary depending on the groups they contain and these groups affect each other's activities. Silyl ethers
(-OTBDMS and -OTMS) and -OH and -Br groups in the skeletal structure of isoindole-1,3(2H)-dione can be
regarded as anticancer agents. In this sense, compounds 13 and 16, containing both silyl ether (-OTBDMS) and -
Br groups, may be regarded as alternative chemotherapeutic drugs. This work may lead to the synthesis of new
isoindole-1,3(2H)-dione compounds containing different silyl ether groups and studies evaluating their
anticancer activities or other biological properties.
Collapse
Affiliation(s)
- Ayse Tan
- Vocational School of Technical Sciences, Mus Alparslan University, Mus 49250, Turkey
| | - Serap Kizilkaya
- Department of Chemistry, Faculty of Arts and Sciences, Mus Alparslan University, Mus 49250, Turkey
| | | | - Atilla Akdemir
- Computer-Aided Drug Discovery Laboratory, Department of Pharmacology, Faculty of Pharmacy, Bezmialem Vakif University, Istanbul 34093, Turkey
| | - Yunus Kara
- Department of Chemistry, Faculty of Sciences, Ataturk University, Erzurum 25240, Turkey
| |
Collapse
|
|
4
|
Pan MS, Cao J, Fan YZ. Insight into norcantharidin, a small-molecule synthetic compound with potential multi-target anticancer activities. Chin Med 2020; 15:55. [PMID: 32514288 PMCID: PMC7260769 DOI: 10.1186/s13020-020-00338-6] [Citation(s) in RCA: 30] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2020] [Accepted: 05/25/2020] [Indexed: 02/07/2023] Open
Abstract
Norcantharidin (NCTD) is a demethylated derivative of cantharidin, which is an anticancer active ingredient of traditional Chinese medicine, and is currently used clinically as a routine anti-cancer drug in China. Clarifying the anticancer effect and molecular mechanism of NCTD is critical for its clinical application. Here, we summarized the physiological, chemical, pharmacokinetic characteristics and clinical applications of NCTD. Besides, we mainly focus on its potential multi-target anticancer activities and underlying mechanisms, and discuss the problems existing in clinical application and scientific research of NCTD, so as to provide a potential anticancer therapeutic agent for human malignant tumors.
Collapse
Affiliation(s)
- Mu-Su Pan
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, 389 Xincun Road, Shanghai, 200065 People’s Republic of China
| | - Jin Cao
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, 389 Xincun Road, Shanghai, 200065 People’s Republic of China
| | - Yue-Zu Fan
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, 389 Xincun Road, Shanghai, 200065 People’s Republic of China
| |
Collapse
|
|
5
|
Zeng L, Liu Y, Pan J, Liu X. Formulation and evaluation of norcanthridin nanoemulsions against the Plutella xylostella (Lepidotera: Plutellidae). BMC Biotechnol 2019; 19:16. [PMID: 30871528 PMCID: PMC6419361 DOI: 10.1186/s12896-019-0508-8] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/16/2018] [Accepted: 03/04/2019] [Indexed: 02/06/2023] Open
Abstract
Background Norcantharidin (NCTD), a demethylated derivative of cantharidin (defensive toxin of blister beetles), has been reported to exhibit insecticidal activity against various types of agricultural pests. However, NCTD applications are limited by its poor water solubility and high dosage requirement. Nanoemulsions have attracted much attentions due to the transparent or translucence appearance, physical stability, high bioavailability and non-irritant in nature. In general, nanoemulsions with small droplet size can enhance the bioavailability of drugs, whereas this phenomenon is likely system dependent. In present study, NCTD nanoemulsions were developed and optimized to evaluate and improve the insecticidal activity of NCTD against Plutella xylostella (Lepidotera: Plutellidae) by a spontaneous emulsification method. Results Triacetin, Cremophor EL and butanol were selected as the constituents of NCTD nanoemulsions via solubility determination, emulsification efficiency and ternary phase diagram construction. Insecticidal activity of NCTD nanoemulsion was associated with the content of surfactant and cosurfactant: (1) Higher effective toxicity exhibited at Smix (surfactant to cosurfactant mass ratio) = 3:1 that may be associated with the changes in interfacial tension; (2) NCTD nanoemulsion at 3:7 < SOR (surfactant to oil mass ratio) < 6:4 was more effective at lower surfactant level, which was attributed to the relatively slow diffusion rate of NCTD hindering by excess surfactant. Interestingly, nanoemulsions with smaller droplets were not found to be more effective in our study. Conclusions The optimized NCTD nanoemulsion (triacetin/Cremophor EL/butanol (60/20/20, w/w)) exhibited effective insecticidal activity (LC50 60.414 mg/l, LC90 185.530 mg/l, 48 h) than the NCTD acetone solution (LC50 175.602 mg/L, LC90 303.050 mg/L, 48 h). Spontaneous emulsifying nanoemulsion employed to formulate this poor water-soluble pesticide is a potential system for agriculture application. Electronic supplementary material The online version of this article (10.1186/s12896-019-0508-8) contains supplementary material, which is available to authorized users.
Collapse
Affiliation(s)
- Liya Zeng
- Key Laboratory of Comprehensive Utilization of Advantage Plants Resources in Hunan South, College of Chemistry and Bioengineering, Hunan University of Science and Engineering, Yongzhou, Hunan, China
| | - Yongchang Liu
- Key Laboratory of Comprehensive Utilization of Advantage Plants Resources in Hunan South, College of Chemistry and Bioengineering, Hunan University of Science and Engineering, Yongzhou, Hunan, China
| | - Jun Pan
- Key Laboratory of Comprehensive Utilization of Advantage Plants Resources in Hunan South, College of Chemistry and Bioengineering, Hunan University of Science and Engineering, Yongzhou, Hunan, China
| | - Xiaowen Liu
- Key Laboratory of Comprehensive Utilization of Advantage Plants Resources in Hunan South, College of Chemistry and Bioengineering, Hunan University of Science and Engineering, Yongzhou, Hunan, China.
| |
Collapse
|
|
6
|
Zhang QY, Yue XQ, Jiang YP, Han T, Xin HL. FAM46C is critical for the anti-proliferation and pro-apoptotic effects of norcantharidin in hepatocellular carcinoma cells. Sci Rep 2017; 7:396. [PMID: 28341836 PMCID: PMC5428258 DOI: 10.1038/s41598-017-00313-6] [Citation(s) in RCA: 36] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/22/2016] [Accepted: 02/20/2017] [Indexed: 02/08/2023] Open
Abstract
Norcantharidin (NCTD), a demethylated analog of cantharidin derived from Chinese traditional medicine blister beetle, has been currently used as an anticancer drug for various cancers including hepatocellular carcinoma (HCC). In this study, for a more comprehensive understanding of the targets of NCTD in HCC, next-generation RNA-Seq was utilized. We revealed that the expression of FAM46C, which has been reported as a tumor suppressor for multiple myeloma, was enhanced after NCTD treatment. Re-analysis of TCGA (The Cancer Genome Atlas) LIHC (liver hepatocellular carcinoma) dataset demonstrated that FAM46C expression was significantly lower in HCC tissues than in normal liver tissues. NCTD injection or FAM46C overexpression could mitigate diethylnitrosamine (DEN)-initiated HCC in mice. Ectopic expression of FAM46C in two HCC cell lines, SMCC-7721 and SK-Hep-1, significantly repressed cell proliferation, and increased cells population in G2/M phase and cell apoptotic rate. We also found that FAM46C overexpression caused a notable decrease in Ras expression, MEK1/2 phosphorylation and ERK1/2 phosphorylation. More importantly, FAM46C knockdown significantly weakened the biological effects of NCTD on HCC cells, which suggested NCTD exerted the anticancer functions partially through up-regulating FAM46C. In conclusion, FAM46C, a tumor suppressor for HCC, is important for the anti-proliferation and proapoptotic effects of NCTD.
Collapse
Affiliation(s)
- Qiao-Yan Zhang
- Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, 200433, P. R. China
| | - Xiao-Qiang Yue
- Department of Traditional Chinese Medicine, Changzheng Hospital, Second Military Medical University, Shanghai, 200433, P. R. China
| | - Yi-Ping Jiang
- Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, 200433, P. R. China
| | - Ting Han
- Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, 200433, P. R. China
| | - Hai-Liang Xin
- Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, 200433, P. R. China.
| |
Collapse
|
|
7
|
Qiu P, Wang S, Liu M, Ma H, Zeng X, Zhang M, Xu L, Cui Y, Xu H, Tang Y, He Y, Zhang L. Norcantharidin Inhibits cell growth by suppressing the expression and phosphorylation of both EGFR and c-Met in human colon cancer cells. BMC Cancer 2017; 17:55. [PMID: 28086832 PMCID: PMC5237309 DOI: 10.1186/s12885-016-3039-x] [Citation(s) in RCA: 34] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/23/2016] [Accepted: 12/23/2016] [Indexed: 11/10/2022] Open
Abstract
BACKGROUND Norcantharidin (NCTD) is a Chinese FDA approved, chemically synthesized drug for cancer treatment. The effect of NCTD on signaling proteins of EGFR and c-Met was systematically elucidated in current study. METHODS Two human colon cancer cell lines, HCT116 and HT29, were used as model systems to investigate the anti-cancer molecular mechanism of NCTD. Cell cycle arrest and early/late apoptosis were analyzed by flow cytometry. The levels of EGFR, phospho-EGFR, c-Met, phospho-c-Met and other related proteins were quantified by western blot analysis. RESULTS NCTD induced cell cycle arrest at G2/M phase in both cell lines. The early and late apoptosis was also observed. Further investigation indicated that NCTD suppressed not only the expression of the total EGFR and the phosphorylated EGFR but also the expression of the total c-Met and the phosphorylated c-Met in colon cancer cells. Moreover, EGFR expression could be mostly restored by co-treatment with MG132, a proteasome inhibitor. In addition, NCTD-induced cell death was comparable to that of the anti-cancer drug gefitinib, a tyrosine kinase inhibitor for EGFR, based on the immunoblot analysis of the expressed proteins after the drug treatment. CONCLUSIONS NCTD might be a useful and inexpensive drug candidate to substitute for gefitinib to serve the treatment needs of cancer patients.
Collapse
Affiliation(s)
- Peiju Qiu
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Siwen Wang
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Ming Liu
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - He Ma
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Xuan Zeng
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Meng Zhang
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Lingling Xu
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Yidi Cui
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Huixin Xu
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Yang Tang
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Yanli He
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China
| | - Lijuan Zhang
- Key Laboratory of Marine Drugs, Chinese Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao, 266003, China. .,Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University, Qingdao, 266003, China.
| |
Collapse
|
|
8
|
Wang W, Deng L, Tang S, Qian Q. Synthesis of Pyrazole-linked Norcantharidin Analogues of Substituted Chromones. J Heterocycl Chem 2016. [DOI: 10.1002/jhet.1665] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Affiliation(s)
- Wei Wang
- Chemistry and Chemical Engineering Institute; Shaoxing University; Shaoxing Zhejiang 312000 People's Republic of China
- Zhejiang Supor Pharmaceuticals; Shaoxing 312000 People's Republic of China
| | - Liping Deng
- Chemistry and Chemical Engineering Institute; Shaoxing University; Shaoxing Zhejiang 312000 People's Republic of China
| | - Shenlong Tang
- Chemistry and Chemical Engineering Institute; Shaoxing University; Shaoxing Zhejiang 312000 People's Republic of China
| | - Qing Qian
- Chemistry and Chemical Engineering Institute; Shaoxing University; Shaoxing Zhejiang 312000 People's Republic of China
| |
Collapse
|
|
9
|
Han HW, Qiu HY, Hu C, Sun WX, Yang RW, Qi JL, Wang XM, Lu GH, Yang YH. Design, synthesis and anti-cancer activity evaluation of podophyllotoxin-norcantharidin hybrid drugs. Bioorg Med Chem Lett 2016; 26:3237-3242. [DOI: 10.1016/j.bmcl.2016.05.063] [Citation(s) in RCA: 21] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2016] [Revised: 05/14/2016] [Accepted: 05/21/2016] [Indexed: 11/26/2022]
|
|
10
|
Yang PY, Hu DN, Kao YH, Lin IC, Chou CY, Wu YC. Norcantharidin induces apoptosis in human prostate cancer cells through both intrinsic and extrinsic pathways. Pharmacol Rep 2016; 68:874-80. [PMID: 27351942 DOI: 10.1016/j.pharep.2016.04.010] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/29/2016] [Revised: 04/12/2016] [Accepted: 04/15/2016] [Indexed: 12/21/2022]
Abstract
BACKGROUND Norcantharidin, a modified pure compound from blister beetles, was previously demonstrated to induce apoptosis of cancer cells. This study investigated its anti-cancer activity in prostate cancer cells and the mechanisms involved. METHODS Two human prostate cancer cell lines, 22Rv1 and Du145, were treated with norcantharidin at concentrations ranging from 3 to 30μg/ml. Cytotoxic effect of norcantharidin was determined by use of the 3-(4,5-dimethylthiazol-yl)-diphenyl tetrazoliumbromide (MTT) assay. The effects of apoptosis were evaluated by cell death assay, Caspase-3, -8, -9 activity and cytochrome c release. The apoptotic related protein expressions (Bcl-2 family and inhibitor of apoptosis proteins) were determined using western blotting. RESULTS An MTT assay revealed that norcantharidin induced cytotoxicity against both prostate cancer cells in dose- and time-dependent manners. Treatment with norcantharidin at 3μg/ml or higher significantly increased oligonucleosomal formation with concomitant appearance of PARP cleavage, implicating the induction of apoptosis. Norcantharidin intrinsically elevated cytosolic cytochrome c levels and activated caspase-3, -8, and -9. Extrinsically, it upregulated the expression of not only the death receptors Fas and DR5 in 22Rv1 cells, but also of RIP and TRADD adaptor proteins in Du145 cells. Mechanistically, norcantharidin increased ratios of pro-/anti-apoptotic proteins and decreased expression of IAP family member proteins, including cIAP1 and survivin, regardless of the distinct status of androgen receptor expression in both cells. CONCLUSIONS Norcantharidin exhibited cytotoxicity against 22Rv1 and Du145 prostate cancer cells by inducing both intrinsic and extrinsic apoptotic pathways and could thus potentially be a remedy for prostate cancer.
Collapse
Affiliation(s)
- Pei-Yu Yang
- Department of Medical Research, Show Chwan Memorial Hospital, Changhua, Taiwan, ROC.
| | - Dan-Ning Hu
- Tissue Culture Center, New York Eye and Ear Infirmary of Mount Sinai, New York, NY, USA.
| | - Ying-Hsien Kao
- Department of Medical Research, E-DA Hospital, Kaohsiung, Taiwan, ROC.
| | - I-Ching Lin
- Department of Family Medicine, Changhua Christian Hospital, Changhua, Taiwan, ROC; School of Medicine, Chung Shan Medical University, Taichung, Taiwan, ROC; School of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC.
| | - Chih-Yuan Chou
- Division of Urology, Department of Surgery, Show-Chwan Memorial Hospital, Changhua, Taiwan, ROC.
| | - Yang-Chang Wu
- School of Pharmacy, College of Pharmacy, China Medical University, Taichung, Taiwan, ROC.
| |
Collapse
|
|
11
|
Abstract
The natural phosphoprotein phosphatase inhibitor cantharidin, primarily used for topical treatment of warts, has later been shown to trigger tumor cell apoptosis and is thus considered for the treatment of malignancy. Similar to apoptosis of tumor cells, erythrocytes may undergo eryptosis, a suicidal cell death characterized by cell shrinkage and translocation of cell membrane phosphatidylserine to the erythrocyte surface. Signaling of eryptosis includes increase of cytosolic Ca2+-activity ([Ca2+]i), ceramide, oxidative stress and dysregulation of several kinases. Phosphatidylserine abundance at the erythrocyte surface was quantified utilizing annexin-V-binding, cell volume from forward scatter, [Ca2+]i from Fluo3-fluorescence, ceramide from antibody binding, and reactive oxidant species (ROS) from 2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence. A 48 h treatment of human erythrocytes with cantharidin significantly increased the percentage of annexin-V-binding cells (≥10 μg/mL), significantly decreased forward scatter (≥25 μg/mL), significantly increased [Ca2+]i (≥25 μg/mL), but did not significantly modify ceramide abundance or ROS. The up-regulation of annexin-V-binding following cantharidin treatment was not significantly blunted by removal of extracellular Ca2+ but was abolished by kinase inhibitor staurosporine (1 μM) and slightly decreased by p38 inhibitor skepinone (2 μM). Exposure of erythrocytes to cantharidin triggers suicidal erythrocyte death with erythrocyte shrinkage and erythrocyte membrane scrambling, an effect sensitive to kinase inhibitors staurosporine and skepinone.
Collapse
|
|
12
|
Wang H, Sun W, Zhang WZ, Ge CY, Zhang JT, Liu ZY, Fan YZ. Inhibition of tumor vasculogenic mimicry and prolongation of host survival in highly aggressive gallbladder cancers by norcantharidin via blocking the ephrin type a receptor 2/focal adhesion kinase/paxillin signaling pathway. PLoS One 2014; 9:e96982. [PMID: 24811250 PMCID: PMC4014585 DOI: 10.1371/journal.pone.0096982] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/18/2013] [Accepted: 04/14/2014] [Indexed: 01/15/2023] Open
Abstract
Vasculogenic mimicry (VM) is a newly-defined tumor microcirculation pattern in highly aggressive malignant tumors. We recently reported tumor growth and VM formation of gallbladder cancers through the contribution of the ephrin type a receptor 2 (EphA2)/focal adhesion kinase (FAK)/Paxillin signaling pathways. In this study, we further investigated the anti-VM activity of norcantharidin (NCTD) as a VM inhibitor for gallbladder cancers and the underlying mechanisms. In vivo and in vitro experiments to determine the effects of NCTD on tumor growth, host survival, VM formation of GBC-SD nude mouse xenografts, and vasculogenic-like networks, malignant phenotypes i.e., proliferation, apoptosis, invasion and migration of GBC-SD cells. Expression of VM signaling-related markers EphA2, FAK and Paxillin in vivo and in vitro were examined by immunofluorescence, western blotting and real-time polymerase chain reaction (RT-PCR), respectively. The results showed that after treatment with NCTD, GBC-SD cells were unable to form VM structures when injecting into nude mouse, growth of the xenograft was inhibited and these observations were confirmed by facts that VM formation by three-dimensional (3-D) matrix, proliferation, apoptosis, invasion, migration of GBC-SD cells were affected; and survival time of the xenograft mice was prolonged. Furthermore, expression of EphA2, FAK and Paxillin proteins/mRNAs of the xenografts was downregulated. Thus, we concluded that NCTD has potential anti-VM activity against human gallbladder cancers; one of the underlying mechanisms may be via blocking the EphA2/FAK/Paxillin signaling pathway.
Collapse
Affiliation(s)
- Hui Wang
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China
| | - Wei Sun
- Department of Surgery, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China
| | - Wen-Zhong Zhang
- Department of Surgery, Shanghai Pudong New Area People's Hospital, Shanghai, P.R. China
| | - Chun-Yan Ge
- Department of Oncology, Shanghai Yangpu Geriatric Hospital, Shanghai, P.R. China
| | - Jing-Tao Zhang
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China
| | - Zhong-Yan Liu
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China
| | - Yue-Zu Fan
- Department of Surgery, Tongji Hospital, Tongji University School of Medicine, Tongji University, Shanghai, P.R. China
| |
Collapse
|
|
13
|
Shen B, He PJ, Shao CL. Norcantharidin induced DU145 cell apoptosis through ROS-mediated mitochondrial dysfunction and energy depletion. PLoS One 2013; 8:e84610. [PMID: 24367681 PMCID: PMC3868658 DOI: 10.1371/journal.pone.0084610] [Citation(s) in RCA: 48] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/30/2013] [Accepted: 11/15/2013] [Indexed: 11/19/2022] Open
Abstract
Norcantharidin (NCTD), a demethylated analog of cantharidin derived from blister beetles, has attracted considerable attentions in recent years due to their definitely toxic properties and the noteworthy advantages in stimulating bone marrow and increasing the peripheral leukocytes. Hence, it is worth studying the anti-tumor effect of NCTD on human prostate cancer cells DU145. It was found that after the treatment of NCTD with different concentrations (25-100 μM), the cell proliferation was significantly inhibited, which led to the appearance of micronucleus (MN). Moreover, the cells could be killed in a dose-/ time-dependent manner along with the reduction of PCNA (proliferating cell nuclear antigen) expression, destruction of mitochondrial membrane potential (MMP), down-regulation of MnSOD, induction of ROS, depletion of ATP, and activation of AMPK (Adenosine 5‘-monophosphate -activated protein kinase) . In addition, a remarkable release of cytochrome c was found in the cells exposed to 100 μM NCTD and exogenous SOD-PEG could eliminate the generation of NCTD-induced MN. In conclusion, our studies indicated that NCTD could induce the collapse of MMP and mitochondria dysfunction. Accumulation of intercellular ROS could eventually switch on the apoptotic pathway by causing DNA damage and depleting ATP.
Collapse
Affiliation(s)
- Bo Shen
- Institute of Radiation Medicine, Fudan University, Shanghai, China
- * E-mail: (C-LS); (BS)
| | - Pei-Jie He
- Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China
| | - Chun-Lin Shao
- Institute of Radiation Medicine, Fudan University, Shanghai, China
- * E-mail: (C-LS); (BS)
| |
Collapse
|
|
14
|
Lee YC, Lee LM, Yang CH, Lin AMY, Huang YC, Hsu CC, Chen MS, Chi CW, Yin PH, Kuo CD, Liao JF, Lee HC. Norcantharidin suppresses cell growth and migration with enhanced anticancer activity of gefitinib and cisplatin in human non-small cell lung cancer cells. Oncol Rep 2013; 29:237-243. [PMID: 23128522 DOI: 10.3892/or.2012.2118] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/17/2012] [Accepted: 09/07/2012] [Indexed: 11/06/2022] Open
Abstract
Norcantharidin is the demethylated analog of cantharidin isolated from blister beetles (Mylabris phalerata Pall.). In this study, we evaluated whether norcantharidin exhibits anticancer effects against the human non-small cell lung cancer cell lines A549 (epidermal growth factor receptor (EGFR) mutation-negative) and PC9 (EGFR mutation-positive). Our results revealed that norcantharidin dose-dependently retards cell growth, arrests cell cycle at G2/M phase, reduces cell migration, and even induces apoptosis at the concentration of 100 µM. Moreover, we found that norcantharidin enhances the anticancer effects of gefitinib and cisplatin. Norcantharidin exhibited similar potency of anticancer effects against the two cell lines with different EGFR mutation status and did not affect EGF-induced EGFR phosphorylation, suggesting that the EGFR signaling may not be the target of norcantharidin. In conclusion, our results suggest that norcantharidin exhibits anticancer effects against non-small cell lung cancer cells in vitro and support its potential as a chemotherapeutic agent for treating non-small cell lung cancer.
Collapse
Affiliation(s)
- Ya-Chun Lee
- Department and Institute of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC
| | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
15
|
Yeh CH, Yang YY, Huang YF, Chow KC, Chen MF. Induction of apoptosis in human Hep3B hepatoma cells by norcantharidin through a p53 independent pathway via TRAIL/DR5 signal transduction. Chin J Integr Med 2012; 18:676-82. [DOI: 10.1007/s11655-012-1206-8] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2010] [Indexed: 11/30/2022]
|
|
16
|
Liu D, Shi P, Yin X, Chen Z, Zhang X. Effect of norcantharidin on the human breast cancer Bcap-37 cells. Connect Tissue Res 2012; 53:508-12. [PMID: 22606958 DOI: 10.3109/03008207.2012.694928] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/03/2023]
Abstract
Norcantharidin (NCTD), a chemically modified form of cantharidin, is a potential anticancer drug. In this study, the effects of NCTD on the cellular viability, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and DNA damage in the human breast cancer cell line Bcap-37 were investigated with confocal and fluorescence microscopy. The cell cycle was further analyzed using the CellQuest software of a Becton-Dickinson FACS flow cytometer. The results indicated that the cellular viability was decreased with the growing concentrations of NCTD and time exposure. Moreover, the fluorescence intensity of ROS was increased, whereas the MMP was decreased in Bcap-37 cells with the growing concentrations of NCTD. NCTD induced a dose-dependent DNA damage and reduced the G1 peak in Bcap-37 cells. The G2/M peak of Bcap-37 was also decreased by the higher concentration of NCTD.
Collapse
Affiliation(s)
- Dongwu Liu
- School of Life Sciences, Shandong University of Technology, Zibo, China
| | | | | | | | | |
Collapse
|
|
17
|
Ding XY, Hong CJ, Liu Y, Gu ZL, Xing KL, Zhu AJ, Chen WL, Shi LS, Zhang XN, Zhang Q. Pharmacokinetics, tissue distribution, and metabolites of a polyvinylpyrrolidone-coated norcantharidin chitosan nanoparticle formulation in rats and mice, using LC-MS/MS. Int J Nanomedicine 2012; 7:1723-35. [PMID: 22619523 PMCID: PMC3356170 DOI: 10.2147/ijn.s29696] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/29/2022] Open
Abstract
A novel formulation containing polyvinylpyrrolidone (PVP) K30-coated norcantharidin (NCTD) chitosan nanoparticles (PVP–NCTD–NPs) was prepared by ionic gelation between chitosan and sodium tripolyphosphate. The average particle size of the PVP–NCTD–NPs produced was 140.03 ± 6.23 nm; entrapment efficiency was 56.33% ± 1.41%; and drug-loading efficiency was 8.38% ± 0.56%. The surface morphology of NCTD nanoparticles (NPs) coated with PVP K30 was characterized using various analytical techniques, including X-ray diffraction and atomic force microscopy. NCTD and its metabolites were analyzed using a sensitive and specific liquid chromatography-tandem mass spectrometry method with samples from mice and rats. The results indicated the importance of the PVP coating in controlling the shape and improving the entrapment efficiency of the NPs. Pharmacokinetic profiles of the NCTD group and PVP–NCTD–NP group, after oral and intravenous administration in rats, revealed that relative bioavailabilities were 173.3% and 325.5%, respectively. The elimination half-life increased, and there was an obvious decrease in clearance. The tissue distribution of NCTD in mice after the intravenous administration of both formulations was investigated. The drug was not quantifiable at 6 hours in all tissues except for the liver and kidneys. The distribution of the drug in the liver and bile was notably improved in the PVP–NCTD–NP group. The metabolites and excretion properties of NCTD were investigated by analyzing rat feces and urine samples, collected after oral administration. A prototype drug and two metabolites were found in the feces, and seven metabolites in the urine. The primary elimination route of NCTD was via the urine. The quantity of the parent drug eliminated in the feces of the PVP–NCTD–NP group, was 32 times greater than that of the NCTD group, indicating that the NPs dramatically increased the reduction quantity from liver to bile. We conclude that PVP–NCTD–NPs are an adequate formulation for enhancing the absorption of NCTD, and significantly improving therapeutic effects targeting the hepatic system. Decarboxylation and hydroxylation were the dominant metabolic pathways for NCTD. Metabolites were mainly excreted into rat kidney and finally into urine.
Collapse
Affiliation(s)
- Xin-Yuan Ding
- Department of Pharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou, People’s Republic of China
| | | | | | | | | | | | | | | | | | | |
Collapse
|
|
18
|
Lu K, Cao M, Mao W, Sun X, Tang J, Shen Y, Sui M. Targeted acid-labile conjugates of norcantharidin for cancer chemotherapy. ACTA ACUST UNITED AC 2012. [DOI: 10.1039/c2jm33069e] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/21/2022]
|
|
19
|
Dong X, Li JC, Jiang YY, Xia MY, Tashiro SI, Onodera S, Ikejima T. p38-NF-κB-promoted mitochondria-associated apoptosis and G2/M cell cycle arrest in norcantharidin-treated HeLa cells. JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH 2012; 14:1008-1019. [PMID: 23281704 DOI: 10.1080/10286020.2012.693481] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/01/2023]
Abstract
Previous study proved that norcantharidin (NCTD) could exert its anticancer activity in a variety of malignant cell lines, including human cervical carcinoma HeLa cells. In this study, we found that NCTD-activated p38 mitogen-activated protein kinase (p38 MAPK)-nuclear transcription factor kappa B (NF-κB) signaling pathway induced mitochondrial apoptotic pathway activation and G2/M cell cycle arrest in HeLa cells. NCTD-induced mitochondria-associated apoptosis was concomitant with the collapse of mitochondrial membrane potential (ΔΨ(m)), translocation of Bax, down-regulation of Bcl-2 expression, and release of cytochrome c. NCTD-led G2/M cell-cycle arrest was associated with the up-regulated p21 and p-cdc25c expression and the down-regulated cyclin B and cdc2 expression. Treatment of the cells with p38 inhibitor SB203580 and NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) showed that p38 functioned upstream of NF-κB, while augmented apoptosis and cell cycle arrest were induced in response to NCTD with NF-κB activation. Intriguingly, NF-κB had a negative feedback regulatory effect on p38 activation. Moreover, NCTD-induced apoptosis and cell cycle arrest were significantly blocked by SB203580 and PDTC but not by pifithrin-α (p53 inhibitor). Therefore, p38-NF-κB induced mitochondrial apoptotic pathway and G2/M cell cycle arrest in NCTD-treated HeLa cells.
Collapse
Affiliation(s)
- Xiu Dong
- China-Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016, China
| | | | | | | | | | | | | |
Collapse
|
|
20
|
Robertson MJ, Gordon CP, Gilbert J, McCluskey A, Sakoff JA. Norcantharimide analogues possessing terminal phosphate esters and their anti-cancer activity. Bioorg Med Chem 2011; 19:5734-41. [DOI: 10.1016/j.bmc.2011.01.031] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/30/2010] [Revised: 01/12/2011] [Accepted: 01/16/2011] [Indexed: 10/18/2022]
|
|
21
|
Yang PY, Chen MF, Kao YH, Hu DN, Chang FR, Wu YC. Norcantharidin induces apoptosis of breast cancer cells: Involvement of activities of mitogen activated protein kinases and signal transducers and activators of transcription. Toxicol In Vitro 2011; 25:699-707. [DOI: 10.1016/j.tiv.2011.01.011] [Citation(s) in RCA: 32] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/21/2010] [Revised: 12/09/2010] [Accepted: 01/18/2011] [Indexed: 01/05/2023]
|
|
22
|
Norcantharidin induces cell cycle arrest and inhibits progression of human leukemic Jurkat T cells through mitogen-activated protein kinase-mediated regulation of interleukin-2 production. Toxicol In Vitro 2011; 25:206-12. [DOI: 10.1016/j.tiv.2010.11.001] [Citation(s) in RCA: 33] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/21/2010] [Revised: 10/06/2010] [Accepted: 11/03/2010] [Indexed: 01/01/2023]
|
|
23
|
Chang C, Zhu YQ, Mei JJ, Liu SQ, Luo J. Involvement of mitochondrial pathway in NCTD-induced cytotoxicity in human hepG2 cells. JOURNAL OF EXPERIMENTAL & CLINICAL CANCER RESEARCH : CR 2010; 29:145. [PMID: 21059274 PMCID: PMC2987898 DOI: 10.1186/1756-9966-29-145] [Citation(s) in RCA: 46] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 05/30/2010] [Accepted: 11/09/2010] [Indexed: 12/13/2022]
Abstract
BACKGROUND Norcantharidin, the demethylated analog of cantharidin derived from a traditional Chinese medicine, Mylabris, has been used in the treatment of anti-cancer effects. However, the detailed mechanisms underlying this process are generally unclear. The aim of this study was to investigate the mechanism of NCTD-induced apoptosis in HepG2 cells. METHODS The cytotoxicity was measured by MTT assay for cellular viability and by flow cytometry. The mitochondrial membrane potential and reactive oxygen species production was evaluated by flow cytometry analysis. The role of caspase activities were assayed using caspase apoptosis detection kit . Western blot analysis was used to evaluate the level of Cyto-C, Bcl-2, Bax, Bid, caspase 3, -9, -8 and PARP expression RESULTS After treatment with NCTD, a decrease in the viability of HepG2 cells and increase in apoptosis were observed. NCTD-induced apoptosis was accompanied by an increase in ROS production, loss of mitochondrial membrane potential and release of cytochrome c(cyto-c) from the mitochondria to the cytosol and down-regulation of anti-apoptotic protein Bcl-2 levels with concurrent up-regulation in pro-apoptotic protein Bax levels. However, another pro-apoptotic molecule, Bid, showed no change in such same treatment. NCTD-increased activity of caspase 9,caspase 3 and the subsequent cleavage caspase substrate PARP were also observed. The expression levels of pro-caspase-8 were not changed after NCTD treatment. CONCLUSION These results indicate that NCTD induced cytotoxicity in HepG2 cells by apoptosis, which is mediated through ROS generation and mitochondrial pathway.
Collapse
Affiliation(s)
- Cheng Chang
- Department of Gastroenterology, Zhongnan Hospital of Wuhan University, Wuhan, 430071, China
| | | | | | | | | |
Collapse
|
|
24
|
Involvement of caspase and MAPK activities in norcantharidin-induced colorectal cancer cell apoptosis. Toxicol In Vitro 2010; 24:766-75. [DOI: 10.1016/j.tiv.2009.12.025] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2009] [Revised: 12/17/2009] [Accepted: 12/21/2009] [Indexed: 01/12/2023]
|
|
25
|
Chang C, Zhu Y, Tang X, Tao W. The anti-proliferative effects of norcantharidin on human HepG2 cells in cell culture. Mol Biol Rep 2010; 38:163-9. [PMID: 20333548 DOI: 10.1007/s11033-010-0090-6] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2009] [Accepted: 03/15/2010] [Indexed: 12/17/2022]
Abstract
Many lines of evidence have shown that Chinese medicine contains many chemical compounds with anticancer effects. Therefore, we tested whether the active ingredients of blister beetles have a therapeutic effect on hepatoma. The aim of this study was to investigate the inhibitive effects of norcantharidin which is extracted from blister beetles on human hepatoma cells HepG2 in vitro and its anticancer mechanism.MTT assay, agarose gel electrophoresis and flow cytometry were used to evaluate HepG2 cells proliferation and apoptosis. The role of caspase activities were assayed using caspase apoptosis detection kit. Western blot analysis was used to evaluate the level of Bcl-2/Bax expression. Our results indicate that norcantharidin inhibited HepG2 cell growth in a time- and dose-dependent manner by MTT assay. HepG2 cells treated with norcantharidin showed typical characteristics of apoptosis including the DNA fragmentation. The activities of caspase-3, -9 were up-regulated after norcantharidin treatment. By western blot analysis, we found the level of Bcl-2 were down-regulated, whereas, the level of Bcl-2 Up-regulated.so we suggest that up-regulation of mitochondrial Bax expression and down-regulation of Bcl-2 expression participated in the apoptosis induced by NCTD. These results suggest that norcantharidin triggers apoptosis in hepato cancer cell lines via the activation of the caspses, mitochondrial pathways, and that this agent may be useful for developing new therapeutic regimens for the treatment of colorectal carcinoma.
Collapse
Affiliation(s)
- Cheng Chang
- Department of Gastroenterology, Zhongnan Hospital of Wuhan University, East-Lake Road 169, 430071, Wuhan, Hubei Province, China
| | | | | | | |
Collapse
|
|
26
|
Yang H, Guo W, Xu B, Li M, Cui J. Anticancer activity and mechanisms of norcantharidin-Nd3II on hepatoma. Anticancer Drugs 2007; 18:1133-7. [PMID: 17893513 DOI: 10.1097/cad.0b013e3282eeb1c5] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
Norcantharidin (NCTD), a demethylated form of cantharidin, is currently used as an anticancer drug in China, but five newly synthesized derivatives have not been tested for antitumor efficacy. In this study, we investigated the in-vitro and in-vivo activity of five derivatives on Bel-7402, HeLa and PC-3M1E8 cell lines on a sulfarhodamine B assay. All of the derivatives showed significant antiproliferative activity, hence we elected to study further one of them, NCTD-Nd3II, in an in-vivo mouse model, and to examine its effects on cell cycle and protein expression. NCTD-Nd3II inhibited H22 tumors in mice in a dose-dependent manner with low toxicity. Flow cytometry results showed that apoptosis and G2/M cell cycle arrest contributed to the cytotoxic and cytostatic effects of NCTD-Nd3II. Further studies showed that Bax and p21 protein expression was upregulated, whereas cyclin B1, Cdc-2 and Bcl-2 protein expression was downregulated. Our findings show that NCTD-Nd3II might be a promising chemotherapeutic agent for hepatomas.
Collapse
Affiliation(s)
- Huayu Yang
- State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center, Beijing, PRC
| | | | | | | | | |
Collapse
|
|
27
|
Rauh R, Kahl S, Boechzelt H, Bauer R, Kaina B, Efferth T. Molecular biology of cantharidin in cancer cells. Chin Med 2007; 2:8. [PMID: 17610718 PMCID: PMC1934358 DOI: 10.1186/1749-8546-2-8] [Citation(s) in RCA: 63] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/02/2007] [Accepted: 07/04/2007] [Indexed: 01/08/2023] Open
Abstract
Herbal medicine is one of the forms of traditional medical practice. Traditional Chinese medicine (TCM) and traditional Vietnamese medicine (TVM) are well-known for their long-standing tradition of herbal medicine. Secreted by many species of blister beetle, most notably by the 'Spanish fly' (Lytta vesicatoria), cantharidin inhibits protein phosphatases 1 and 2A (PP1, PP2A). Blister beetle has been used in Asian traditional medicine to treat Molluscum contagiosum virus (MCV) infections and associated warts, and is now also used for cancer treatment. A combination of both genomic and postgenomic techniques was used in our studies to identify candidate genes affecting sensitivity or resistance to cantharidin. Cantharidin was not found to be related to multidrug resistance phenotype, suggesting its potential usefulness for the treatment of refractory tumors. Oxidative stress response genes diminish the activity of cantharidin by inducing DNA strand breaks which may be subject to base excision repair and induce apoptosis in a p53- and Bcl2-dependent manner. Cantharidin is one of many natural products used in traditional Chinese medicine and traditional Vietnamese medicine for cancer treatment. Combined methods of pharmaceutical biology and molecular biology can help elucidate modes of action of these natural products.
Collapse
Affiliation(s)
- Rolf Rauh
- State of Maryland Department of Health and Mental Hygiene, Maryland, USA
| | - Stefan Kahl
- Institute of Pharmaceutical Sciences, University of Graz, Graz, Austria
| | | | - Rudolf Bauer
- Institute of Pharmaceutical Sciences, University of Graz, Graz, Austria
| | - Bernd Kaina
- Institute of Toxicology, University of Mainz, Mainz, Germany
| | - Thomas Efferth
- Pharmaceutical Biology (C015), German Cancer Research Centre, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany
| |
Collapse
|
|
28
|
Kok SHL, Chui CH, Lam WS, Chen J, Lau FY, Wong RSM, Cheng GYM, Lai PBS, Leung TWT, Yu MWY, Tang JCO, Chan ASC. Synthesis and structure evaluation of a novel cantharimide and its cytotoxicity on SK-Hep-1 hepatoma cells. Bioorg Med Chem Lett 2006; 17:1155-9. [PMID: 17240140 DOI: 10.1016/j.bmcl.2006.12.039] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2006] [Revised: 11/22/2006] [Accepted: 12/12/2006] [Indexed: 11/30/2022]
Abstract
A remarkable control of the potency of cantharimide is described based on the electronic properties of functional group and it exhibits a relatively less toxic effect to the non-malignant hematological disorder bone marrow cells.
Collapse
Affiliation(s)
- Stanton Hon Lung Kok
- Lo Ka Chung Centre for Natural Anti-Cancer Drug Development, Institute of Molecular Technology for Drug Discovery and Synthesis, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, PR China
| | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
29
|
Fan YZ, Fu JY, Zhao ZM, Chen CQ. Effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells. World J Gastroenterol 2005; 11:2431-7. [PMID: 15832413 PMCID: PMC4305630 DOI: 10.3748/wjg.v11.i16.2431] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate the effect of norcantharidin on proliferation and invasion of human gallbladder carcinoma GBC-SD cells in vitro and its anticancer mechanism.
METHODS: Human gallbladder carcinoma GBC-SD cells were cultured by cell culture technique. The growth and the invasiveness of GBC-SD cells in vitro were evaluated by the tetrazolium-based colorimetric assay and by the Matrigel experiment and the crossing-river test. Expression of PCNA, Ki-67, MMP2 and TIMP2 proteins of GBC-SD cells was determined by streptavidin–biotin complex method.
RESULTS: In vitro norcantharidin inhibited the growth and proliferation of GBC-SD cells in a dose- and time-dependent manner, with the IC50 value of 56.18 μg/mL at 48 h. Norcantharidin began to inhibit the invasion of GBC-SD cells at the concentration of 5 μg/mL, and the invasive action of GBC-SD cells was inhibited completely and their crossing-river time was prolonged significantly at 40 μg/mL. After treatment with norcantharidin, the expression of PCNA, Ki-67, and MMP2 was significantly decreased. With the increase in TIMP2 expression, the MMP2 to TIMP2 ratio was decreased significantly (P<0.05).
CONCLUSION: Norcantharidin inhibits the proliferation and growth of human gallbladder carcinoma cells in vitro at relatively low concentrations by inhibiting PCNA and Ki-67 expression. Its anti-invasive activity may be the result of decrease in MMP2 to TIMP2 ratio and reduced cell motility.
Collapse
Affiliation(s)
- Yue-Zu Fan
- Department of General Surgery, Tongji Hospital of Tongji University, 389 Xincun Road, Shanghai 200065, China.
| | | | | | | |
Collapse
|
|
30
|
Efferth T, Rauh R, Kahl S, Tomicic M, Böchzelt H, Tome ME, Briehl MM, Bauer R, Kaina B. Molecular modes of action of cantharidin in tumor cells. Biochem Pharmacol 2005; 69:811-8. [PMID: 15710358 DOI: 10.1016/j.bcp.2004.12.003] [Citation(s) in RCA: 60] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/30/2004] [Accepted: 12/03/2004] [Indexed: 10/25/2022]
Abstract
Cancer chemotherapy is often limited by patient's toxicity and tumor drug resistance indicating that new drug development and modification of existing drugs is critical for improving the therapeutic response. Traditional Chinese medicine is a rich source of potential anticancer agents. In particular, cantharidin (CAN), the active principle ingredient from the blister beetle, Mylabris, has anti-tumor activity, but the cytotoxic mechanism is unknown. In leukemia cells, cantharidin induces apoptosis by a p53-dependent mechanism. Cantharidin causes both DNA single- and double-strand breaks. Colony-forming assays with knockout and transfectant cells lines showed that DNA polymerase beta, but not ERCC1, conferred increased cell survival after cantharidin treatment, indicating that base excision repair (BER), rather than nucleotide excision repair (NER), is important for CAN-induced DNA lesions. Oxidative stress-resistant thymic lymphoma-derived WEHI7.2 variants are also more resistant to cantharidin. These data suggest that cantharidin treatment causes oxidative stress that provokes DNA damage and p53-dependent apoptosis.
Collapse
Affiliation(s)
- Thomas Efferth
- German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
| | | | | | | | | | | | | | | | | |
Collapse
|
|
31
|
Kok SH, Cheng SJ, Hong CY, Lee JJ, Lin SK, Kuo YS, Chiang CP, Kuo MYP. Norcantharidin-induced apoptosis in oral cancer cells is associated with an increase of proapoptotic to antiapoptotic protein ratio. Cancer Lett 2005; 217:43-52. [PMID: 15596295 DOI: 10.1016/j.canlet.2004.07.045] [Citation(s) in RCA: 81] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/19/2004] [Revised: 07/05/2004] [Accepted: 07/15/2004] [Indexed: 11/30/2022]
Abstract
Norcantharidin (NCTD), the demethylated analogue of cantharidin, has been used to treat human cancers in China since 1984. It was recently found to be capable of inducing apoptosis in human colon carcinoma, hepatoma and glioblastoma cells by way of an elusive mechanism. In this study, we demonstrated that NCTD also induces apoptosis in human oral cancer cell lines SAS (p53 wild-type phenotype) and Ca9-22 (p53 mutant) as evidenced by nuclear condensation, TUNEL labeling, DNA fragmentation and cleavage of PARP. Apoptosis induced by NCTD was both dose- and time-dependent. We found NCTD did not induce Fas and FasL, implying that it activated other apoptosis pathways. Our data showed that NCTD caused accumulation of cytosolic cytochrome c and activation of caspase-9, suggesting that apoptosis occurred via the mitochondria mediated pathway. NCTD enhanced the expression of Bax in SAS cells consistent with their p53 status. Moreover, we showed that NCTD downregulated the expression of Bcl-2 in Ca9-22 and Bcl-XL in SAS. Our results suggest that NCTD-induced apoptosis in oral cancer cells may be mediated by an increase in the ratios of proapoptotic to antiapoptotic proteins. Since oral cancer cells with mutant p53 or elevated Bcl-XL levels showed resistance to multiple chemotherapeutic agents, NCTD may overcome the chemoresistance of these cells and provide potential new avenues for treatment.
Collapse
Affiliation(s)
- Sang-Heng Kok
- School of Dentistry, College of Medicine, National Taiwan University, 1 Chang-Te Street, Taipei 10016, Taiwan, ROC
| | | | | | | | | | | | | | | |
Collapse
|
|
32
|
An WW, Wang MW, Tashiro SI, Onodera S, Ikejima T. Norcantharidin induces human melanoma A375-S2 cell apoptosis through mitochondrial and caspase pathways. J Korean Med Sci 2004; 19:560-6. [PMID: 15308848 PMCID: PMC2816891 DOI: 10.3346/jkms.2004.19.4.560] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/20/2022] Open
Abstract
Norcantharidin (NCTD) is the demethylated form of cantharidin, which is the active substance of mylabris. To examine the pathway of NCTD-induced A375-S2 cell death, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-dipheyltetrazolium bromide (MTT) assay, photomicroscopical observation, DNA agarose gel electrophoresis, caspase activity assay and Western blot analysis were carried out. A375-S2 cells treated with NCTD exhibited several typical characteristics of apoptosis. The inhibitory effect of NCTD on human melanoma, A375-S2 cells, was partially reversed by the inhibitors of pan-caspase, caspase-3 and caspase-9. The activities of caspase-3 and -9 were significantly increased after treatment with NCTD at different time. The expression of inhibitor of caspase-activated DNase was decreased in a time-dependent manner, simultaneously, the ratio of Bcl-2/Bax or Bcl-xL/Bax was decreased and the expression ratio of proteins could be reversed by caspase-3 inhibitor. The expression of cytochrome c in cytosol was increased after NCTD treatment and caspase- 3 inhibitor had no significant effect on the up-regulation of cytochrom c. These results suggest that NCTD induced A375-S2 cell apoptosis and the activation of caspase and mitochondrial pathway were involved in the process of NCTD-induced A375-S2 cell apoptosis.
Collapse
Affiliation(s)
- Wei-Wei An
- China-Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang, China
| | | | | | | | | |
Collapse
|
|
33
|
Chen YN, Cheng CC, Chen JC, Tsauer W, Hsu SL. Norcantharidin-induced apoptosis is via the extracellular signal-regulated kinase and c-Jun-NH2-terminal kinase signaling pathways in human hepatoma HepG2 cells. Br J Pharmacol 2003; 140:461-70. [PMID: 12970086 PMCID: PMC1574052 DOI: 10.1038/sj.bjp.0705461] [Citation(s) in RCA: 49] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022] Open
Abstract
Norcantharidin (NCTD) is an anticancer drug routinely used against hepatoma in China. Previously, we reported that NCTD could induce mitotic arrest and apoptosis in human hepatoma HepG2 cells. However, the intracellular signaling pathways involved in NCTD-induced apoptotic cell death are still obscure. Caspase inhibitors were used to clarify the role of specific caspase in NCTD-triggered apoptotic process. Results showed that activation of caspase-9/caspase-3 cascade is required for NCTD-induced apoptotic death. To decipher the upstream signals for NCTD-induced apoptosis, we characterized the involvement of mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38MAPK. The role of their downstream targets, transcription factors activating protein-1 (AP-1), and nuclear factor kappaB (NF-kappaB) in NCTD-induced apoptosis was also analyzed. Immunoblot analyses and in vitro kinase assay demonstrated that NCTD-induced apoptosis was accompanied by the elevations of the levels of phosphorylated form and kinase activity of ERK and JNK, but not p38MAPK. The inhibitor of ERK pathway (U0126 or PD98059) or JNK pathway (SP600125) markedly prevented kinase activation, and also greatly reduced NCTD-induced apoptotic cell death. Increased DNA-binding activity of AP-1 and NF-kappaB was also observed after NCTD treatment. Inhibition of NF-kappaB activation by PDTC or inhibition of AP-1 activation by curcumin drastically blocked NCTD-induced cell death. These results imply that activation of ERK and JNK, and modulation of downstream transcription factors NF-kappaB and AP-1, may be involved in NCTD-induced apoptosis.
Collapse
Affiliation(s)
- Yan-Nian Chen
- School of Chinese Medicine, China Medical College, Taichung, Taiwan
- China Medical College Peikang Hospital, Yunlin, Taiwan
- St. Martin De Porres Hospital, Chaiyi, Taiwan
| | - Chi-Chih Cheng
- Department of Education & Research, Taichung Veterans General Hospital, Taichung, Taiwan
| | - Jung-Chou Chen
- China Medical College Research Institute of Chinese Medicine, Taiwan
- Show Chwan Memorial Hospital, Department of Chinese Medicine, Taiwan
| | - Wei Tsauer
- School of Post Baccalaureate Chinese Medicine Clinical Medical College, Taiwan
| | - Shih-Lan Hsu
- School of Chinese Medicine, China Medical College, Taichung, Taiwan
- Department of Education & Research, Taichung Veterans General Hospital, Taichung, Taiwan
- Author for correspondence:
| |
Collapse
|
|
34
|
Huh JE, Kang KS, Ahn KS, Kim DH, Saiki I, Kim SH. Mylabris phalerlata induces apoptosis by caspase activation following cytochrome c release and Bid cleavage. Life Sci 2003; 73:2249-62. [PMID: 12927594 DOI: 10.1016/s0024-3205(03)00568-x] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/10/2023]
Abstract
Mylabris phalerata (MP) is an insect that has been used for the treatment of cancer in oriental medicine. In the present study, the butanol (BuOH) fraction of MP (BFMP) was examined to determine whether it can exert anti-cancer activity through an apoptotic pathway with little toxicity. BFMP was found to have a specific cytotoxic effect on human monocytic leukemic U937 cells (IC(50) = 140 microg/ml) rather than on peripheral blood mononuclear lymphocytes (PBML, IC(50) = over 500 microg/ml). BFMP also induced the morphological changes of apoptosis, such as chromatin condensation, cell shrinking and DNA fragmentation at a concentration of 31.25 microg/ml. In addition, BFMP significantly increased the portion of apoptotic annexin-V positive cells in a dose-dependent manner, and effectively activated caspases (cysteine aspartase) cascade involving caspases 8, 9 and 3. BFMP also effectively cleaved Bid, a death agonist member of the Bcl-2 family and (poly(ADP-ribose)polymerase) (PARP) and induced the subsequent release of cytochrome c from mitochondria into the cytosol. However, it did not affect Bcl-2 and Bax expression. Taken together, these data suggest that the BuOH extract of Mylabris phalerata can induce apoptosis in U937 cells by caspase cascade activation in conjunction with cytochrome c release, induced by a product of Bid. Therefore, we conclude that BFMP has anti-cancer activity, which is achieved through apoptosis and is associated with little toxicity.
Collapse
Affiliation(s)
- Jeong-Eun Huh
- Department of Oncology, Graduate School of East-West Medicine, KyungHee University, 1 Seochunri, Yongin 449-701, South Korea
| | | | | | | | | | | |
Collapse
|