【摘要】 目的研究肿瘤特异性凋亡蛋白基因(apoptin)在诱导人宫颈癌细胞凋亡过程中信号转导机制。方法用含有apoptin基因的真核表达载体瞬间转染体外培养的人宫颈癌细胞(hela);采用逆转录-聚合酶链反应(RT-PCR)、DNA凝胶电泳、流式细胞术检测hela细胞的凋亡;以比色法检测胱天蛋白酶-8(caspase-8)和胱天蛋白酶-3(caspase-3)的相对活性。结果 Apoptin基因瞬间转染的hela细胞可出现典型的细胞凋亡所具有的DNA梯状带;流式细胞术检测发现,实验组细胞凋亡率〔(40.45±0.76)%〕明显高于正常对照组〔(3.25±0.16)%〕和载体对照组〔(3.55±0.12)%〕(P<0.01);caspase-3的活性升高,但caspase-8活性无明显变化。结论 Apoptin基因可通过激活caspase-3诱导hela细胞凋亡。更多还原

【Abstract】 Objective To study whether apoptin gene induces apoptosis in hela cells via the activation caspase-8 and caspase-3.Methods The hela cells were transiently transfected with recombinant pcDNAA3 plasmid,which contained apoptin gene and the apoptosis was measured with DNA agarose electrophoresis,RT-PCR and flow cytometry.Caspase-8 and caspase-3 relative activity was analyzed with colorimetric assay at various time.Results The apoptin gene could induce apoptosis of hela cells in vitro and the mRNA of apoptin gene could be detected by RT-PCR.The activity of caspase-3 increased in hela cells after transfected with apoptin gene,but that of caspase-8 did not change.Conclusion Apoptin gene can induce apoptosis in hela cells via activ ation of caspase-3.更多还原