共济失调毛细血管扩张突变基因沉默可抑制人乳腺癌细胞的迁移和侵袭  

Ataxia-telangiectasia mutated gene-silencing inhibits the migration and invasion of human breast cancer cells

作　　者：郎磊[1] 杜燕娥[1] 席磊[1] 周明莉[2] 孙可馨 阴嘉莉[1] 陈燕林[1] 柳满然[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016 [2]成都市第三人民医院检验科,四川成都610031

出　　处：《肿瘤》2016年第10期1090-1097,共8页Tumor

基　　金：国家自然科学基金面上项目(编号:81472476)~~

摘　　要：目的 :探讨共济失调毛细血管扩张突变(ataxia-telangiectasia mutated,ATM)基因沉默对人三阴性乳腺癌MDA-MB-231细胞增殖、迁移和侵袭的影响。方法 :将携带ATM-sh RNA及其阴性对照序列(作为阴性对照组)的重组慢病毒载体分别感染人三阴性乳腺癌MDA-MB-231细胞,获得ATM基因稳定沉默及其对照细胞株;同时,设置未经病毒感染的空白对照组细胞。经嘌呤霉素筛选后,荧光显微镜下观察各组细胞的感染效率,并采用实时荧光定量PCR和蛋白质印迹法分别检测ATM mRNA及蛋白的表达水平。采用MTT法、FCM法、细胞划痕愈合实验和Transwell小室实验分别检测ATM基因沉默对MDA-MB-231细胞增殖、周期分布、迁移和侵袭能力的影响。结果:成功构建了稳定沉默ATM基因的人三阴性乳腺癌MDA-MB-231细胞株。与空白对照组和阴性对照组相比,ATM基因沉默组细胞的增殖能力和细胞周期分布均无明显变化(P值均>0.05),但细胞迁移和侵袭能力均明显减弱(P值均<0.05)。结论:在人三阴性乳腺癌MDA-MB-231细胞中,ATM表达下调可明显抑制细胞的迁移和侵袭。Objective： To investigate the effects of ataxia-telangiectasia mutated（ATM） gene-silencing on the proliferation, migration and invasion of human triple-negative breast cancer MDA-MB-231 cells. Methods： The recombinant lentiviral vectors with ATM gene-targetedspecific sh RNA or the negative control sequence（as the negative control group） were infected into the human triple-negative breast cancer MDA-MB-231 cells to obtain the ATM gene-silencing cells. At the same time, the uninfected MDA-MB-231 cells was used as the blank control group. After screening by puromycin, the infection efficiency of each group was observed under a fluorescence microscope. The expressions of ATM mRNA and protein in MDA-MB-231 cells in the three groups were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The effects of ATM gene-silencing on proliferation, cycle distribution, migration and invasion of MDA-MB-231 cells were analyzed by MTT method, FCM, cell wound healing assay and Transwell assays, respectively.Results： The human triple-negative breast cancer MDA-MB-231 cells with stable ATM genesilencing were established successfully. Compared with the blank control and negative control groups, the proliferation and cell cycle distribution of MDA-MB-231 cells in ATM gene-silencing group had no significant change（all P〈0.05）, but the migration and invasion abilities of MDA-MB-231 cells in ATM gene-silencing group were decreased significantly（all P〈0.05）.Conclusion： The down-regulation of ATM gene expression can significantly inhibit the migration and invasion of human triple-negative breast cancer MDA-MB-231 cells.

关 键 词：乳腺肿瘤 共济失调毛细血管扩张突变蛋白质类 细胞增殖 细胞迁移分析 肿瘤浸润 

分 类 号：R737.9[医药卫生—肿瘤]