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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 effect 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 vector carrying ATM-sh RNA and its negative control sequence (negative control group) was used to infect human triple-negative breast cancer MDA-MB-231 cells respectively to obtain stable ATM gene silencing and its control cell line. , Set the control group of cells without virus infection. After puromycin screening, the infection efficiency of cells in each group was observed under a fluorescence microscope. The expression of ATM mRNA and protein was detected by real-time fluorescence quantitative PCR and Western blot respectively. The effects of ATM gene silencing on the proliferation, cell cycle distribution, migration and invasion of MDA-MB-231 cells were detected by MTT assay, FCM assay, cell scratch healing assay and Transwell chamber assay. Results: The human triple negative breast cancer MDA-MB-231 cell line was successfully constructed with stable silenced ATM gene. Compared with the blank control group and negative control group, the proliferation and cell cycle distribution of ATM gene silencing group had no significant changes (P> 0.05), but the cell migration and invasion ability were significantly decreased (P <0.05 ). Conclusion: Down-regulation of ATM expression in human triple-negative breast cancer MDA-MB-231 cells can significantly inhibit cell migration and invasion.