钩藤酸E通过线粒体途径诱导HepG2细胞凋亡

Mitochondria-dependent Mechanisms Involved in HepG2 cell Apoptosis Induced by Uncarinic Acid E

目的研究钩藤酸E诱导人肝癌HepG2细胞凋亡的机制。方法采用MTT法分析确定钩藤酸E对肿瘤细胞及对小鼠骨髓体外增殖抑制作用,显微镜观察分析细胞凋亡的形态学变化,乳酸脱氢酶测定分析细胞死亡机制,免疫印迹实验检测线粒体相关蛋白的表达。结果钩藤酸E可明显地抑制HepG2细胞生长,且呈明显的量效关系和时效关系,而对小鼠骨髓细胞无抑制作用。Hoechst33258荧光染色可见凋亡小体产生,乳酸脱氢酶分析证明钩藤酸E引起细胞死亡为凋亡和坏死同时进行。免疫印迹结果显示上调的Bax和下调的Bcl-2和Bcl-xL表达证实了钩藤酸E诱导的HepG2细胞凋亡是通过线粒体途径发挥作用,并导致细胞色素C的释放。结论钩藤酸E可明显地诱导HepG2细胞凋亡,并经历了线粒体信号转导途径。

Aim To study the mechanisms of uncarinic acid E-induced HepG2 cell apoptosis. Methods Cytotoxicity assay by MTT, observasion of morphology, LDH release and Western blot analysis were carried out. Results Uncarinic acid E inhibited HepG2 cell growth in a dose- and time-dependent manner, and did not show inhibitory action on marrow in mice in vitro. Apoptotic bodies were observed in Hoechst33258 fluorescein stain. LDH assay proved that apoptosis and necrosis undertook simultaneously in death of HepG2 cell treated with uncarinic acid E. Up-regulation of Bax and down-regulation of Bcl-2 and Bcl-xL expression contributed to uncarinic acid E induced HepG2 cell apoptosis, and led to release of cytochrome c. Conclusion Mitochondria-dependent signal transduction pathways are involved in uncarinic acid E-induced HepG2 cell apoptosis.