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目的研究caspase-3在肿瘤特异性凋亡基因诱导人黑色素瘤细胞A375凋亡中的作用。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黑色瘤细胞A375;采用RT-PCR、DNA凝胶电泳、流式细胞术检测A375细胞的凋亡;以比色法检测caspase-3的相对活性。结果Ap-optin基因瞬间转染的A375细胞可出现典型的细胞凋亡所具有的DNA梯状带;流式细胞术发现实验组细胞凋亡率明显高于其他各组(P<0.01);转染后24h实验组caspase-3的活性开始升高,72h达高峰,明显高于其他各组(P<0.01)。结论Apoptin基因可通过激活caspase-3诱导人黑色素瘤细胞A375凋亡。
Objective To investigate the role of caspase-3 in the apoptosis of human melanoma cell line A375 induced by tumor-specific gene. Methods The human melanoma A375 cells were transiently transfected with the eukaryotic expression vector apoptin in vitro. The apoptosis of A375 cells was detected by RT-PCR, DNA gel electrophoresis and flow cytometry. The expressions of caspase- 3 relative activity. Results Apoptosis DNA ladder was found in A375 cells transiently transfected with Ap-optin gene. Flow cytometry showed that apoptosis rate of cells in experimental group was significantly higher than that in other groups (P <0.01) The activity of caspase-3 began to increase in experimental group 24 h after staining, reaching the peak at 72 h, which was significantly higher than that of other groups (P <0.01). Conclusion Apoptin can induce A375 apoptosis in human melanoma cells by activating caspase-3.