RNA干扰iNOS基因对颊鳞癌细胞凋亡影响的实验研究

Study on the Influence on Apoptosisy of Bca885 Cell by RNA Interference to iNOS

目的研究RNA干扰iNOS基因的表达对肿瘤细胞凋亡的影响。方法采用RNA干扰技术,通过脂质体介导,将含有iNOS特异性发夹状小RNA(short hairpin RNA,shRNA)的pGenesil-1质粒转染入颊鳞癌Bca885细胞株。观察pGenesil-1质粒载体转染率、免疫组化检测细胞中iNOS的蛋白表达的改变情况、流式细胞仪检测Bca885细胞的凋亡率。采用SPSS12.0统计软件统计分析。结果在脂质体介导下,将含iNOS特异性shRNA的质粒转染入颊鳞癌细胞后,实验组Bca885细胞中iNOS蛋白表达低于两对照组,具有统计学意义(P<0.01);三组细胞的凋亡率分别为12.05±0.18、3.31±0.12和2.46±0.09,实验组高于两对照组,具有统计学上意义(P<0.01)。结论RNA干扰可以降低iNOS基因的表达,达到基因沉默的效果;沉默iNOS基因可提高Bca885细胞的凋亡,从而降低Bca885细胞的细胞增殖活性。

Objective To investigate the availability of RNA technique and the relation between iNOS and apoptosis. Methods The vector that involved short hairpin RNA（shRNA） ofiNOS was transfected to Bca885 cell by LIPOFECTAMINE 2000. And at the same time, negative control and blank control were set up. The change of iNOS expression was observed using immunohistochemistry technique, the apoptosis rate was examined by flow cytomtery. SPSS 12.0 package was used in statistic analysis. The difference between groups was con- sidered significant when P〈0.05. Results The expression ofiNOS in Psilencer-iNOS group cell was higher than that in control groups （P〈0. 01）; The apoptosis rate of three groups Bca885 cells were 12.05 ± 0.18, 3.31 ± 0.12 and 2.46 ± 0.09 （P〈0.01）. Conclusion Downexpression ofiNOS by RNAi can promot apoptosis of Bca885 cells and has an anti-proliferation activity effect.