Induction of Apoptosis in Human Hep3B Hepatoma Cells by Norcantharidin through a p53 Independent Pathway via TRAIL/DR5 Signal Transduction

• Published in: Chinese journal of integrative medicine Vol. 18; no. 9; pp. 676 - 682
• Main Author: 叶宗勋 杨玉燕 黄雅芳 周宽基 陈明丰
• Format: Journal Article
• Language: English
• Published: Heidelberg Chinese Association of Traditional and Western Medicine 01.09.2012
• Subjects: 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; apoptosis; norcantharidin; caspase; death receptors
• ISSN: 1672-0415; 1993-0402; 1993-0402
• DOI: 10.1007/s11655-012-1206-8

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,