细胞色素C在apoptin诱导宫颈癌Hela细胞凋亡中的作用

Effect of cytochrome C on apoptosis of Hela cells induced by apoptin

目的研究肿瘤特异性凋亡基因(apoptin)在诱导Hela细胞凋亡中的信号转导机制。方法用含有apoptin基因的真核表达载体瞬间转染体外培养的Hela细胞;采用MTT法检测Hela细胞的凋亡;以比色法检测caspase-8和caspase-3的相对活性;Western blotting检测凋亡细胞中细胞色素C的表达量。结果 MTT法证明ap-optin基因瞬间转染的Hela细胞凋亡率明显高于其他各组(P<0.01);caspase-3的活性升高,但caspase-8活性没有明显变化;细胞色素C释放量明显增多。结论 Apoptin基因可能通过促进线粒体释放细胞色素C激活caspase-3,进而诱导Hela细胞凋亡。

Objective To study whether apoptin gene induces apoptosis of Hela cells via the release of cytochrome C from mitochondria and the activation caspase-8 and caspase-3.Method The Hela cells were transiently transfected with recombinant pcDNAA3 plasmid,which contained apoptin gene,and the apoptosis were measured with MTT methods. Caspase-8 and caspase-3 relative activity were analyzed by colorimetric assay at various time. The change of cytochrome C was monitored with Western blotting.Result The apoptin gene could induce apoptosis of Hela cells in vitro and the activity of caspase-3 began to rise in Hela cells after transfected with apoptin gene,but that of caspase-8 did not change. Western blotting showed more expression of cytochrome C in transfected Hela cells than that in control. Conclusion Apoptin gene can induce apoptosis in Hela cells via promoting the release of cytochrome C from mitochondria and then active caspase-3.