肿瘤特异性凋亡基因；细胞；凋亡；半胱天冬蛋白酶_

Apoptin基因诱导A375细胞凋亡信号转导机制

http://www.100md.com 《中国公共卫生》 2007年第4期

肿瘤特异性凋亡基因；细胞；凋亡；半胱天冬蛋白酶,,肿瘤特异性凋亡基因；细胞；凋亡；半胱天冬蛋白酶,1材料与方法,2结果,3讨论,参考文献

     摘要：目的研究肿瘤特异性凋亡基因(apoptin)在诱导人黑色素瘤细胞A375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人黑色素瘤细胞A375；采用RT-PCR、DNA凝胶电泳、流式细胞仪分析检测A375的细胞的凋亡；以比色法检测半胱天冬蛋白酶8(caspase-8)和caspase-3的相对活性。结果 Apoptin基因瞬间传染的A375细胞可出现典型的细胞凋亡所具有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)；Caspase-3的活性升高，但caspase-8活性没有明显变化。结论 Apoptin基因可通过激活caspase-3诱导人黑色素瘤细胞A375凋亡。

    关键词：肿瘤特异性凋亡基因；细胞；凋亡；半胱天冬蛋白酶

    Study on signal transduction in apoptosis of human melanoma cells A375 induced by apoptin

    GE Quanzhi,ZHAO Hongli,DONG Ping.

    Lab of Molecular Biology,Center for Disease Control and Prevention,Shenyang Military Command(Shenyang 110034,China)Abstract： Objective To study whether apoptin gene induces apoptosis in human melanoma cells A375 via the activation caspase-8 and caspase-3.Methods The human melanoma cells A375 were transiently transfected with recombinant pcDNAA3 plasmid,which contained apoptin gene,and the apoptosis was measured by DNA agarose electrophoresis,RT-PCR and flow cytometry.Caspase-8 and caspase-3 relative activity was analyzed by colorimetric assay at various time.Results The apoptin gene could induce apoptosis of human melanoma cells A375in vitro and the mRNA of apoptin gene could be detected by RT-PCR.The activity of caspase-3 began to rise in A375 cells after transfected with apoptin gene,but that of caspase-8 did not change.Conclusion Apoptin gene can induce apoptosis in human melanoma cells A375via active caspase-3. ......

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