RNAi抑制Survivin基因的表达对乳腺癌SKBr-3细胞的影响  

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

作　　者：李昆仑[1,2] 崔明[1] 刘嘉[1] 冯雁康[1] 马彦光[1] 朱俊[1] 

机构地区：[1]成都军区昆明总医院普通外科,云南昆明650032 [2]昆明医学院研究生部,云南昆明650031

出　　处：《昆明医学院学报》2012年第2期17-22,共6页Journal of Kunming Medical College

基　　金：云南省自然科学基金资助项目(2010ZC176)

摘　　要：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±0.3)%;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.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（P0.01）,the highest cell inhibition rate reached（43.1±0.3）%.RT-PCR result showed the survivin mRNA expression in survivin-siRNA groups was downregulated（P0.01）,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（P0.01）,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（P0.01）,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.

关 键 词：SURVIVIN RNAI 细胞凋亡 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]