Department of Gastroenterology, Zhongnan Hospital of Wuhan University, East-Lake Road 169, 430071, Wuhan, Hubei Province, China.
Mol Biol Rep. 2011 Jan;38(1):163-9. doi: 10.1007/s11033-010-0090-6. Epub 2010 Mar 24.
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.
大量证据表明,中药含有许多具有抗癌作用的化合物。因此,我们测试了斑蝥素的活性成分是否对肝癌有治疗作用。本研究旨在探讨斑蝥素对体外人肝癌细胞 HepG2 的抑制作用及其抗癌机制。MTT 法、琼脂糖凝胶电泳和流式细胞术用于评估 HepG2 细胞的增殖和凋亡。用 caspase 凋亡检测试剂盒测定 caspase 活性在其中的作用。Western blot 分析用于评估 Bcl-2/Bax 表达水平。我们的结果表明,斑蝥素通过 MTT 法抑制 HepG2 细胞生长,呈时间和剂量依赖性。用斑蝥素处理的 HepG2 细胞表现出典型的凋亡特征,包括 DNA 片段化。用斑蝥素处理后,caspase-3、-9 的活性上调。通过 Western blot 分析,我们发现 Bcl-2 水平下调,而 Bcl-2 水平上调。因此,我们认为线粒体 Bax 表达上调和 Bcl-2 表达下调参与了 NCTD 诱导的凋亡。这些结果表明,斑蝥素通过激活 caspase、线粒体途径诱导肝癌细胞凋亡,该药物可能有助于开发治疗结直肠癌的新治疗方案。