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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 investigate the effect of RNA interference on the apoptosis of A375 cells after inhibiting survivin gene expression in malignant melanoma cell line A375. Methods The siRNA eukaryotic expression vector pU-survivin-siRNA targeting the apoptosis-suppressing gene survivin was constructed and transfected into A375 cells by electroporation. The expression of survivin was detected by Western blotting and apoptosis was detected by flow cytometry. The change. Results After transfection with pU-survivin-siRNA, the expression of survivin in A375 cells (0.24±0.02) was significantly lower than that in the control group (0.98±0.21), and the apoptosis rate of the test group cells was significantly decreased. (83%) increased significantly compared to the control group (28%). Conclusions The expression of survivin can be inhibited by RNA interference and the apoptosis of A375 cells can be increased.