siRNA干扰CHRNA5基因的表达抑制尼古丁对人肺癌细胞的促增殖作用  

siRNA interference of CHRNA5 gene expression inhibits nicotine-induced proliferation of human lung cancer cells

作　　者：赵云[1] 马晓丽[1,2] 汪运山[1] 郏雁飞[1] 张雪萍[2] 郑燕[1] 肖东杰[1] 

机构地区：[1]山东大学附属济南市中心医院中心实验室,山东济南250013 [2]山东师范大学生命科学学院,山东济南250014

出　　处：《中国肿瘤生物治疗杂志》2010年第6期604-608,共5页Chinese Journal of Cancer Biotherapy

基　　金：中国博士后科学基金特别资助(No.200902575);济南留学人员创业计划项目(No.20080407)~~

摘　　要：目的:研究siRNA干扰尼古丁乙酰胆碱受体α5(cholinergic receptor nicotinic alpha5,CHRNA5)基因表达后对尼古丁促人肺癌A549细胞增殖作用的影响。方法:设计并合成CHRNA5-siRNA片段,并经脂质体介导转染A549细胞。荧光定量PCR、Western blotting检测转染后A549细胞中CHRNA5 mRNA和蛋白的表达,MTT法检测CHRNA5-siRNA对尼古丁促A549细胞增殖作用的影响。结果:成功构建了CHRNA5-siRNA并成功转染A549细胞。CHRNA5-siRNA转染组A549细胞的CHRNA5mRNA表达量明显低于si-NC转染对照组(0.196±0.044vs0.944±0.027,P<0.01),CHRNA5-siRNA转染组A549细胞的CHRNA5蛋白表达量明显低于si-NC转染组(0.267±0.031vs0.745±0.035,P<0.01)。在无尼古丁作用时,CHRNA5-siRNA的转染不能抑制A549细胞的增殖。在尼古丁作用下,A549细胞增殖显著增加(P<0.01),CHRNA5-siRNA转染可明显抑制尼古丁的促增殖作用(P<0.01)。结论:siRNA下调肺癌A549细胞中CHRNA5的表达可以抑制尼古丁促细胞增殖作用,CHRNA5可能是肺癌治疗的潜在靶点之一。Objective：To study the effect of siRNA targeting CHRNA5 （cholinergic receptor nicotinic alpha 5） gene expression on nicotine-induced proliferation of lung cancer A549 cells.Methods： CHRNA5-siRNA fragment was designed and synthesized,and then transfected into A549 cells by Lipofectamine assay.Expressions of CHRNA5 mRNA and protein in transfected-A549 cells were examined by FQ-PCR and Western blotting analysis,respectively.The effect of CHRNA5-siRNA on nicotine-induced proliferation of A549 cells was detected by MTT method.Results： CHRNA5-siRNA was successfully constructed and transfected into A549 cells.CHRNA5 mRNA expression in CHRNA5-siRNA-transfected A549 cells was significantly lower than that in control si-NC control siRNA transfected A549 cells （0.196±0.044 vs 0.944±0.027,P0.01）,and CHRNA5 protein expression in CHRNA5-siRNA-transfected A549 cells was also lower than that in si-NC-transfected A549 cells （0.267±0.031 vs 0.745±0.035,P0.01）.When A549 cells were not treated with nicotine,CHRNA5-siRNA transfection inhibited the proliferation of A549 cells.But Nicotine increased the proliferation of A549 cells （P0.01）,and this positive effect was inhibited by CHRNA5-siRNA transfection （P0.01）.Conclusion： siRNA targeting CHRNA5 expression can inhibit nicotine-induced proliferation of A549 cells,which suggests that CHRNA5 may serve as a potential target for gene therapy of lung cancer.

关 键 词：α5-尼古丁乙酰胆碱受体(CHRNA5) SIRNA 肺癌 尼古丁 增殖 

分 类 号：R734.2[医药卫生—肿瘤] R730.54[医药卫生—临床医学]