| 摘要: |
| ［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Survivin基因可能成为乳腺癌基因治疗的一个新靶点. |
| 关键词: ［关键词］Survivin RNAi 细胞凋亡 乳腺癌 |
| DOI： |
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| Effect of Survivin Gene Expression Inhibition by RNAi on Breast Cancer SKBr-3 Cells |
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LI Kun－lun
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| Abstract: |
| ［Abstract］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. |
| Key words: ［Key words］Survivin RNAi Cell apoptosis Breast cancer |