刘霜莉,王强.RNAi技术沉默Survivin基因抑制肺癌细胞株H1299增殖并诱导其凋亡[J].内科急危重症杂志,2015,21(21):
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| DOI： |
| 中文关键词: Survivin siRNA H1299细胞 凋亡 |
| 英文关键词:Survivin RNA interference H1299 cell cell proliferation apoptosis |
| 基金项目:湖北省教育厅科学研究计划重点项目（D20131103） |
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| 中文摘要: |
| 目的 研究RNAi技术沉默Survivin基因对人肺癌细胞株H1299增殖和凋亡的影响。方法 将 Survivin特异性siRNA 表达载体siRNA-Sur转染H1299细胞, 利用倒置荧光显微镜和流式细胞仪检测转染效率,通过Western blot和Realtime-PCR方法检测RNAi沉默Survivin 表达的效果, MTT 法检测RNAi沉默Survivin对H1299细胞增殖的影响,采用流式细胞术分析细胞周期分布和凋亡。结果 siRNA-Sur转染H1299细胞48h后,细胞的Survivin基因表达明显减少,空白对照组Survivin mRNA的相对表达量为1.0,siRNA-空白对照组为0.98,而siRNA-Sur组为0.32(P<0.01);导致H1299细胞G2期阻滞,空白对照组为23.7% ,而siRNA-Sur组为50.1%(P<0.01);细胞抑制率和凋亡率增加, 转染48h,白对照组凋亡率为5.8%,无关对照组为6.2%,而siRNA-Sur组达50.6%(P<0.01)。 结论 RNAi沉默H1299细胞Survivin基因表达抑制细胞增殖,并诱导H1299细胞凋亡。 |
| 英文摘要: |
| Objective To evaluate the effect of Survivin special siRNA on proliferation and apoptosis of human lung cancer H1299 cells. Methods The expression vector of pgsiRNA-sur was constructed and transfected into H1299 cells by Lipofectin. The protein and mRNA expression of Survivin was detected by Western blot and real time PCR.The changes of cell cycle after transfection were examined by flow cytometry and MTT assay, respectively.Results In H1299 cells, the protein and mRNA levels of Survivin were significantly decreased after transfection, and reduction of proliferation was related to an increase in the fraction of G2 phase. Conclusion The Survivin special siRNA silenced Survivin, decreased H1299 cells proliferation and induced their apoptosis. |
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