RNA干扰抑制PC-3细胞survivin基因表达并诱导细胞凋亡的实验研究

RNA Interference Inhibits the Expression of Survivin and Induces the Apoptosis of PC-3 Cells

目的:研究RNA干扰抑制人前列腺癌PC-3细胞survivin mRNA和蛋白表达并诱导细胞凋亡的效果。方法:设计、构建针对survivin基因的RNAi表达载体,分别以质粒A、B组,阴性序列组和空白对照E组用脂质体法转染PC-3细胞,应用RT-PCR及Western印迹、流式细胞术检测其对survivin mRNA和蛋白表达并诱导细胞凋亡的效果。结果:各组细胞survivin蛋白的表达强度分别为(18.94±0.63)%、(16.35±0.23)%、(46.41±0.76)%、(46.20±1.47)%。各组细胞中survivin mRNA的表达强度分别为(27.94±1.43)%、(24.51±1.37)%、(49.46±0.71)%、(48.49±1.32)%。各组细胞凋亡率分别为(12.80±1.33)%、(16.48±1.00)%、(3.03±0.62)%、(2.96±0.41)%。统计学表明构建的两种阳性表达载体均有效抑制了survivin mRNA和蛋白表达并诱导细胞凋亡。结论:RNAi可有效抑制PC-3细胞survivin mRNA和蛋白表达,并诱导细胞凋亡。

Objecive： To investigate the inhibitory effect of RNA interference （RNAi） on the expression of survivin mRNA and its inducibility of the apoptosis of PC-3 cells. Methods ： siRNA expression vectors were designed and constructed to be directed at survivin and transfected into PC-3 cells by Lipofectamine in 4 groups： plasmid A, plasmid B, negative sequence and control E. RT-PCR, Western blot and flow cytometry were used to detect the expression of suvivin mRNA and the apoptosis of PC-3 cells. Results： The expression rates of survivin protein were 18.94% ± 0.63%, 16.35 ± 0.23%, 46.41% ± 0.76% and 46.20 ± 1.47%, those of survivin mRNA were 27.94% ± 1.43%, 24.51% ± 1.37%, 49.46% ±0.71% and 48.49% ± 1.32%, and the apoptosis rates of PC-3 cells were 12.80% ±1.33%, 16.48% ±1.00%, 3.03% ±0.62% and2.96% ±0.41% respectively in different groups. Conclusion ： RNAi can effectively inhibit the expression of survivin mRNA and induce the apoptosis of PC-3 cells.