RNA干扰内源性一氧化氮合酶基因对舌鳞状细胞癌细胞增殖活性影响的实验研究

Influence of iNOS silencing by RNA interference on proliferation activity of Tca8113 cell

目的研究内源性一氧化氮合酶(inducible nitric oxide synthase,iNOS)对肿瘤细胞凋亡和增殖活性的影响。方法采用 RNA 干扰技术,将含有 iNOS 基因的特异性短发夹状 RNA(shorthairpin RNA,shRNA)片断的质粒载体转染入 Tca8113细胞,免疫组化检测细胞中 iNOS 的蛋白表达改变情况;流式细胞仪及甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTY)法检测细胞凋亡率和生长曲线的变化情况。结果质粒转染入 Tca8113细胞后,实验组细胞中 iNOS 蛋白表达显著低于两对照组(P<0.01);而凋亡率明显高于两对照组(P<0.01);生长曲线显示实验组细胞增殖活性受到一定抑制。结论 RNA 干扰技术沉默 iNOS 基因可提高 Tca8113细胞的凋亡,并降低细胞的增殖活性。

Objective To investigate the apoptosis and proliferation activity of Tca8113 cells. Methods The vector that involves short hairpin RNA of iNOS was transfected to Tca8113 cells. The change of iNOS expression was observed using immunohistochemistry technique, the apoptosis rate examined by flow cytomtery, and the proliferation Tca8113 cells examined by methyl thiazolyl tetrazolium （MTT）. Results The expression of iNOS in Psilencer-iNOS group was lower than that in control groups （ P 〈 0. 01 ）, the apoptosis rate was higher than that in control groups（ P 〈 0. 01 ） ; whereas the proliferation activity of Tca8113 cells in Psilencer-iNOS group was lower than that in control groups . Conclusions Down expression of iNOS by RNAi can promots apoptosis of Tca8113 cells and has an anti-proliferation activity effect.