摘要
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 explore the proliferation and apoptosis of human breast cancer MCF-7 cell by applying RNA interference technique to silence survivin gene.Methods：Chemically synthesized small interference RNA targeting Survivin was transfected into human breast cancer cell lines MCF-7 using lipidosome,then observe the corpuscular transfection efficiency by inverted microscope,detect cell survival rate by MTT colorimereic analysis antigenic,detect cell cycle and apoptotic index by flow cytometry（FCM）,observe the restraining efficiency of the protein expression and survivin mRNA by Western Blot.Results：Survivin-siRNA effectively inhibited survivin mRNA expression（P〈0.05） cell,proliferation（P〈0.05）,and induced cell cycle arrest in G2/M phase with a significant decrease of cells in S phase（P〈0.05）.Conclusion：Blocking the expression of survivin with RNA interference technology can significantly suppress proliferation of MCF-7 cell and induce apoptosis to a certain degree.RNAi targeted to survivin has a potential value in gene therapy of breast cancer.
出处
《现代肿瘤医学》
CAS
2009年第3期410-413,共4页
Journal of Modern Oncology
