RNAi抑制Survivin基因的表达对乳腺癌SKBr-3细胞的影响

目的 应用RNA干扰技术抑制人Survivin基因表达,观察其对乳腺癌SKBr-3细胞增殖和凋亡的影响.方法 设计、合成靶向Survivin基因的siRNA基因片段,应用脂质体包埋转染乳腺癌SKBr-3细胞,采用MTT比色法检测细胞增殖率、RT-PCR和Western blot法观察乳腺癌细胞Survivin mRNA和蛋白质的表达、流式细胞仪检测细胞凋亡率.结果 MTt检测显示Survivin-siRNA组对细胞增殖有明显抑制作用(P<0.05),其细胞最高抑制率为(43.1±O.3)%;RT-PCR检测显示Survivin-siRNA组mRNA表达明显下调(P<0.01),其相对表达量分别为0.203 ±0.018、0.229±0.019,抑制率分别为55.2%、49.4%;Western blot检测显示Survivin-siRNA组蛋白表达下调(P<0.01),其蛋白相对表达量分别为0.702±0.007、0.684±0.016,抑制率分别为34.8%、36.4%;流式细胞仪检测显示Survivin-siRNA组细胞凋亡率明显增高(P<0.01),分别为(14.2±1.4)%、(16.8±0.6)%.结论 Survivin-siRNA能有效封闭Survivin的表达,抑制SKBr-3细胞增殖并诱导细胞凋亡,推测Sur-vivin基因可能成为乳腺癌基因治疗的一个新靶点.

Effect of Survivin Gene Expression Inhibition by RNAi on Breast Cancer SKBr-3 Cells

Objective To observe the effect of survivin gene expression inhibition by small RNA interference(siRNA)on the proliferation and apoptosis of the breast cancer SKBr-3 cells.Methods We designed and synthetized siRNA targeted survivin gene,and used liposome embedding method to transfect the siRNA into the breast cancer SKBr-3 cells.Then we used MTT method to detect cells proliferation rate,Western blot and RT-PCR to observe survivin mRNA and protein expression;Flow cytometry(FCM) to test cell apoptosis rate.Results MTT result showed the proliferation of cells in survivin-siRNA group was inhibited obviously(P<0.01),the highest cell inhibition rate reached(43.1±0.3)%.RT-PCR result showed the survivin mRNA expression in survivin-siRNA groups was downregulated(P<0.01),the relative expression of mRNA was 0.203±0.018 and 0.229±0.019,the inhibition ratio was 55.2% and 49.4%,respectively.Western blot result showed the survivin protein expression in survivin-siRNA groups was downregulated(P<0.01),the relative expression was 0.702±0.007 and 0.684±0.016,the inhibition ratio was 34.8% and 36.4%,respectively.Flow cytometry result showed that cell apoptosis rate was increased obviously in survivin-siRNA groups(P<0.01),which was(14.2±1.4)% and(16.8±0.6)%.Conclusions Survivin-siRNA can effectively inhibit survivin gene expression,thus inhibit SKBr-3 cell proliferation and induce apoptosis.Survivin gene may be a new target for breast cancer therapy,which provides theory basis for breast cancer gene therapy.