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目的:探讨RNA干扰技术沉默survivin基因表达对人乳腺癌细胞增殖和凋亡的影响。方法:设计并合成靶向survivin的小分子干扰RNA(siRNA),在脂质体(lipidosome)的介导下转染人乳腺癌细胞株MCF-7,荧光倒置显微镜观察细胞的转染效率;MTT(四甲基偶氮唑盐)比色法分析检测各组癌细胞的存活率;流式细胞计数检测各组癌细胞周期和凋亡指数;Western Blot方法观察对MCF-7细胞survivin mRNA和蛋白质表达的抑制效率。结果:Survivin-siRNA能有效封闭survivin基因的表达,使survivin的mRNA相对水平明显降低(P<0.05);Survivin-siRNA能明显抑制细胞增殖(P<0.05);Survivin-siRNA使细胞G2/M期细胞百分比明显增加,S期的细胞百分比显著减少(P<0.05)。结论:利用RNA干扰技术沉默survivin基因的表达可以显著抑制MCF-7细胞的增殖,并在一定程度上诱导其自发凋亡,靶向survivin的RNA干扰技术在乳腺癌的基因治疗中具有一定的研究价值。
Objective: To investigate the effects of silencing survivin gene expression by RNA interference on the proliferation and apoptosis of human breast cancer cells. Methods: Small interfering RNA (siRNA) targeting survivin was designed and synthesized. The human breast cancer cell line MCF-7 was transfected by lipidosome. The transfection efficiency of the cells was observed by inverted fluorescence microscope; MTT. Colorimetric analysis was used to determine the survival rate of each group of cancer cells; flow cytometry was used to detect the cell cycle and apoptosis index of each group; Western Blot was used to observe the expression of survivin mRNA and protein in MCF-7 cells. The inhibitory efficiency of expression. RESULTS: Survivin-siRNA can effectively block the expression of survivin gene and reduce the relative level of survivin mRNA (P<0.05). Survivin-siRNA can significantly inhibit cell proliferation (P<0.05); Survivin-siRNA can make cell G2/M phase The percentage of cells was significantly increased, and the percentage of cells in S phase was significantly reduced (P<0.05). Conclusion: Silencing the expression of survivin gene by RNA interference can significantly inhibit the proliferation of MCF-7 cells, and to some extent induce its spontaneous apoptosis. RNA interference targeting survivin has certain research in the gene therapy of breast cancer. value.