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基因表达的效果,MTT法检测RNAi沉默Survivin基因对基因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48 h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48 h,空白对照组凋亡率为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。
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 transfected into H1299 cells. The protein and mRNA expression of survivin was detected by Western blot and real time PCR. The changes of cell cycle and cells proliferation were examined by FCM and MTT assay,respectively. Results： In H1299 cells,the protein and mRNA levels of survivin were significantly decreased after transfection,survivin mRNA of control team is 1. 0,siRNA-blank control team is 0. 98,siRNA-Sur team is 0. 32（ P 0. 01）,and reduction of proliferation was related to an increase in the fraction of G2 phase,survivin mRNA of control team is 23. 7%,siRNA-Sur team is 50. 1% after 48 hour of transfection（ P 0. 01）. The ratio of inhibit and apoptosis of H1299 were significantly increased,survivin mRNA of control team is 5. 8%,siRNA-blank control team is 6. 2%,siRNA-Sur team is 50. 6% after 48 hour of transfection（ P 0. 01）. Conclusions： The survivin special siRNA silenced survivin,decreased H1299 cells proliferation and induced apoptosis.
出处
《内科急危重症杂志》
2015年第4期308-311,共4页
Journal of Critical Care In Internal Medicine
基金
湖北省教育厅科学研究计划重点项目(No:D20131103)
