Induction of Apoptosis in Human Hep3B Hepatoma Cells by Norcantharidin through a p53 Independent Pathway via TRAIL/DR5 Signal Transduction
Objective： To investigate the inhibitory activities of norcantharidin （NCTD）, a demethylated analogue of cantharidin, on Hep3B cells （a human hepatoma cell line） with deficiency of p53. Methods： The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of tr...
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| Published in | Chinese journal of integrative medicine Vol. 18; no. 9; pp. 676 - 682 |
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| Main Author | |
| Format | Journal Article |
| Language | English |
| Published |
Heidelberg
Chinese Association of Traditional and Western Medicine
01.09.2012
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1672-0415 1993-0402 1993-0402 |
| DOI | 10.1007/s11655-012-1206-8 |
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| Abstract | Objective： To investigate the inhibitory activities of norcantharidin （NCTD）, a demethylated analogue of cantharidin, on Hep3B cells （a human hepatoma cell line） with deficiency of p53. Methods： The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined. Results： NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G2M phase arrest occursat low concentration （≤25 μ mol/L） but G0G1 phase arrest at high concentration （50 μ mol/L）. The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation. Conclusion： NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G2M or G0G1 phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway, |
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| AbstractList | To investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell line) with deficiency of p53.OBJECTIVETo investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell line) with deficiency of p53.The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined.METHODSThe survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined.NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G(2)M phase arrest occurs at low concentration ([Symbol: see text] 25 μmol/L) but G(0)G(1) phase arrest at high concentration (50 μmol/L). The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation.RESULTSNCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G(2)M phase arrest occurs at low concentration ([Symbol: see text] 25 μmol/L) but G(0)G(1) phase arrest at high concentration (50 μmol/L). The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation.NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G(2)M or G(0)G(1) phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway.CONCLUSIONNCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G(2)M or G(0)G(1) phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway. To investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell line) with deficiency of p53. The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined. NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G(2)M phase arrest occurs at low concentration ([Symbol: see text] 25 μmol/L) but G(0)G(1) phase arrest at high concentration (50 μmol/L). The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation. NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G(2)M or G(0)G(1) phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway. Objective： To investigate the inhibitory activities of norcantharidin （NCTD）, a demethylated analogue of cantharidin, on Hep3B cells （a human hepatoma cell line） with deficiency of p53. Methods： The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined. Results： NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G2M phase arrest occursat low concentration （≤25 μ mol/L） but G0G1 phase arrest at high concentration （50 μ mol/L）. The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation. Conclusion： NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G2M or G0G1 phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway, Objective To investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell line) with deficiency of p53. Methods The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined. Results NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G 2 M phase arrest occurs at low concentration (⩽ 25 μmol/L) but G 0 G 1 phase arrest at high concentration (50 μmol/L). The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation. Conclusion NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G 2 M or G 0 G 1 phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway. |
| Author | 叶宗勋 杨玉燕 黄雅芳 周宽基 陈明丰 |
| AuthorAffiliation | Department of Neurology, Show Chwan Memorial Hospital,Changhua, Taiwan, China Graduate Institute of Life Sciences,National .Chung Hsing. University, Taichung, Taiwan：China; Departmentof Medical Research, Show-Chwan Memorial Hospital, Changhua, Taiwan, China Graduate Institute of Biomedical Sciences, National Chung Hsing University,Taichung, Taiwan, China Department of Gastroenterology and Heaptology, E-DA Hospital, Kaohsiung, Taiwan, China Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan, China |
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| Cites_doi | 10.1007/s00432-002-0326-5 10.1016/j.bcp.2003.12.013 10.1016/0145-2126(91)90163-N 10.4161/cbt.7.2.5335 10.1016/S0168-8278(01)00284-7 10.1016/S0304-3835(98)00251-1 10.1016/j.canlet.2004.07.045 10.1053/jhep.2000.16266 10.1097/00002371-200111000-00003 10.1006/bbrc.2000.3341 10.1016/0959-8049(95)00050-X 10.1096/fj.08-111005 10.1016/0378-8741(89)90062-7 10.1016/0022-1759(86)90215-2 10.1002/hep.510290102 10.1016/S0014-5793(00)01375-2 10.1002/ijc.10479 10.1074/jbc.R800084200 10.1016/S0304-3835(97)00206-1 10.1016/S0006-2952(00)00506-2 10.1042/bse0380009 10.1038/nrc2693 |
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| DocumentTitleAlternate | Induction of Apoptosis in Human Hep3B Hepatoma Cells by Norcantharidin through a p53 Independent Pathway via TRAIL/DR5 Signal Transduction |
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| Keywords | apoptosis norcantharidin caspase death receptors |
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| Notes | 11-4928/R norcantharidin;caspase;apoptosis;death receptors YEH Chung-hsin,YANG Yu-yen,HUANG Ya-fang,CHOW Kuan-chih,and CHEN Ming-feng（ 1. Department of Neurology, Show Chwan Memorial Hospital, Changhua, Taiwan, China; 2. Graduate Institute of Life Sciences, National Chung Hsing University, Taichung, Taiwan, China; 3. Department of Medical Research, Show Chwan Memorial Hospital, Changhua, Taiwan, China; 4. Graduate Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan, China; 5. Department of Gastroenterology and Heaptology, E-DA Hospital, Kaohsiung, Taiwan, China; 6. Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan, China） Objective： To investigate the inhibitory activities of norcantharidin （NCTD）, a demethylated analogue of cantharidin, on Hep3B cells （a human hepatoma cell line） with deficiency of p53. Methods： The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined. Results： NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G2M phase arrest occursat low concentration （≤25 μ mol/L） but G0G1 phase arrest at high concentration （50 μ mol/L）. The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation. Conclusion： NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G2M or G0G1 phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
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| Snippet | Objective： To investigate the inhibitory activities of norcantharidin （NCTD）, a demethylated analogue of cantharidin, on Hep3B cells （a human hepatoma cell... Objective To investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell... To investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell line) with... |
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| SubjectTerms | Antibodies, Neoplasm - pharmacology Antibodies, Neutralizing - pharmacology Apoptosis - drug effects Bridged Bicyclo Compounds, Heterocyclic - pharmacology B细胞 Carcinoma, Hepatocellular - enzymology Carcinoma, Hepatocellular - pathology Caspase 10 - metabolism Caspase 3 - metabolism Caspase Inhibitors - pharmacology caspase-3 Cell Cycle Checkpoints - drug effects Cell Line, Tumor Cell Proliferation - drug effects DNA Fragmentation - drug effects Humans Immunohistochemistry Liver Neoplasms - enzymology Liver Neoplasms - pathology Medicine Medicine & Public Health Original Article p53 Receptors, TNF-Related Apoptosis-Inducing Ligand - metabolism Signal Transduction - drug effects TNF-Related Apoptosis-Inducing Ligand - metabolism Tumor Suppressor Protein p53 - metabolism 信号转导 去甲斑蝥素 时间依赖性 细胞凋亡 诱导 |
| Title | Induction of Apoptosis in Human Hep3B Hepatoma Cells by Norcantharidin through a p53 Independent Pathway via TRAIL/DR5 Signal Transduction |
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