NNK抑制ATM基因表达的实验研究  

Experimental study on the inhibition of ATM expression by NNK in non-small cell lung cancer cells

作　　者：陈俊生 董琪 曹玉祥 吴志浩[5] CHEN Junsheng;DONG Qi;CAO Yuxiang;WU Zhihao(School of Anesthesiology,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]皖南医学院麻醉学院,安徽芜湖241002 [2]皖南医学院,芜湖安徽241002 [3]皖南医学院临床医学院,安徽芜湖241002 [4]安徽师范大学生命科学学院,安徽芜湖241000 [5]皖南医学院细胞生物学教研室,生物活性大分子重点实验室,安徽芜湖241002

出　　处：《皖南医学院学报》2018年第2期106-108,共3页Journal of Wannan Medical College

基　　金：安徽省自然科学基金项目(1708085MH203)

摘　　要：目的:研究烟草致癌物质4-甲基亚硝胺基-1-3-吡啶基-1-丁酮(NNK)在非小细胞肺癌细胞H1299中对ATM基因表达的影响。方法:采用Western Blot的方法检测不同时间段NNK处理的人非小细胞肺癌细胞H1299中ATM蛋白的表达量;构建ATM启动子萤火虫荧光素酶报告基因质粒;用双荧光素酶报告基因系统检测经NNK处理的H1299细胞中ATM启动子的活性。结果:Western Blot证实H1299细胞中,NNK在增加了p-ATM(表明ATM被激活)的同时抑制了ATM蛋白的表达。结论:测序结果显示ATM启动子质粒构建成功;双荧光素酶报告基因检测系统证实了在H1299细胞中NNK抑制ATM启动子的活性,从而抑制ATM基因的表达。Objective:To investigate the effect of 4-methylnitrosamino-1-3-pyridyl-1-butanone(NNK)on ataxia-telangiectasia mutated(ATM)gene expression in non-small cell lung cancer(NSCLC)cell line H1299.Methods:Western blot was performed to detect the expression of ATM in H1299 cells treated with NNK at different time intervals,and plasmid of the firefly luciferase reporter gene was developed by fusing to the ATM promoter to test its activity in NNK-treated H1299 cells that was measured in a dual luciferase reporter gene assay.Results:Western blotting indicated that NNK was capable of increasing phospho-ATM levels and inhibiting the expression of ATM protein in H1299 cells.Conclusion:ATM promoter luciferase reporter plasmid was successfully constructed by sequencing verification.Dual luciferase reporter gene assay confirms that NNK is able to inhibit the ATM promoter activity H1299 cells.

关 键 词：ATM NNK 启动子克隆 DNA损伤 肿瘤 

分 类 号：R734.2[医药卫生—肿瘤] R393[医药卫生—临床医学]