siRNA抑制生存素基因对恶性黑素瘤细胞A375凋亡的影响

Effect of siRNA targeting survivin gene on the apoptosis of malignant melanoma cell line A375

目的探讨RNA干扰技术抑制恶性黑素瘤细胞株A375生存素基因表达后,对A375细胞凋亡的影响。方法构建针对凋亡抑制基因生存素的siRNA真核表达载体pU-生存素-siRNA,用电穿孔法转染A375细胞,采用蛋白质印迹技术检测生存素的表达,并用流式细胞仪检测细胞凋亡的变化。结果转染pU-生存素-siRNA后,生存素在A375细胞中的表达(0．24±0．02)较对照组(0．98±0．21)明显下降, 试验组细胞的凋亡率(83％)较对照组(28％)明显增加。结论通过RNA干扰技术可抑制生存素的表达, 诱导A375细胞的凋亡增加。

Objective To study the effect of siRNA targeting survivin gene on the apoptosls of malignant melanoma cell line A375. Methods The eucaryotic expression vector of pU-survivin-siRNA was constructed and transfected into the A375 cells by electroporation. The protein expression of survivin was examined by Western blotting, and cell apoptosis by flow cytometry. Results The transfection of pU-survivin-siRNA significantly down-regulated survivin expression （ 0.24 ±0.02 in the transfected group versus 0.98 ±0.21 in the control group ） in A375 cells, and promoted cell apoptosis （ 83% in the transfected group versus 28% in the control group, P 〈 0.05 ）. Conclusion The siRNA targeting survivin gene could specially suppress survivin gene expression and induce apoptosis of A375 cells, which may provide an additional option for the treatment of malignant melanoma.