shRNA靶向沉默CNTN1表达抑制乳腺癌细胞株MDA-MB-468增殖及克隆形成能力  

shRNA silencing CNTN1 gene expression inhibits proliferation and clone formation of breast cancer MDA-MB-468 cells

作　　者：贺赛 耿洁 杨晓民 侯艳妮 范拥国 赵静 张静远 陈楠 HE Sai;GENG Jie;YANG Xiaomin;HOU Yanni;FAN Yongguo;ZHAO Jing;ZHANG Jingyuan;CHEN Nan(Department of Breast Oncology,Shaanxi Provincial Cancer Hospital,Shaanxi Xi'an 710061,China;Department of Cardiovascular Internal Medicine,the Second Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Xi'an 710004,China)

机构地区：[1]陕西省肿瘤医院乳腺病院,陕西西安710061 [2]西安交通大学第二附属医院心内科,陕西西安710004

出　　处：《现代肿瘤医学》2021年第4期564-568,共5页Journal of Modern Oncology

基　　金：陕西省自然科学基础研究计划(编号:2020JM-680)。

摘　　要：目的:探讨短发夹RNA(shRNA)靶向沉默CNTN1基因表达对乳腺癌MDA-MB-468细胞增殖及克隆形成能力的影响。方法:采用RT-PCR和Western blot法检测乳腺癌细胞株MCF7-ADR、MDA-MB-468、MCF7以及Hs578T中的CNTN1 mRNA和蛋白表达水平。采用Lipofectamine~(TM)2000向MDA-MB-468细胞株转染成功构建的靶向沉默CNTN1表达的shRNA载体片段,并采用RT-PCR法和Western blot法进行鉴定。分别通过MTT法、流式细胞仪技术和平板克隆实验检测各组细胞增殖及克隆形成能力的改变。结果:CNTN1 mRNA和蛋白表达水平在MDA-MB-468中最高。沉默组MDA-MB-468细胞发生G1期阻滞,增殖能力明显降低(P <0.05),克隆形成能力明显减弱(P <0.05)。结论:CNTN1基因在乳腺癌MDA-MB-468细胞中高表达,沉默其表达可抑制乳腺癌MDA-MB-468细胞增殖和克隆形成能力。Objective:To investigate the effect of short hairpin RNA(shRNA)targeted silencing of CNTN1 gene on proliferation and clone formation of breast cancer MDA-MB-468 cells.Methods:Real-time quantitative PCR(RT-PCR)and Western blot were used to detect the mRNA and protein levels of CNTN1 in MCF7-ADR,MDA-MB-468、MCF7 and Hs578T cells.MDA-MB-468 cells were randomly transfected with the successfully constructed shRNA vector fragments targeting CNTN1 gene(silencing group)or nonsense shRNA fragments(control group)via Lipofectamine TM 2000.CNTN1 mRNA and protein expression was detected after 48 h transfection by RT-PCR and Western blot.The proliferation and clone formation abilities of cells were detected by MTT,clone formation assay and flow cytometry,respectively.Results:The relative expression level of CNTN1 mRNA and protein in MDA-MB-468 cells had the highest expression significantly(P<0.05).MDA-MB-468 cells in transfection group underwent G 1 phase block.The capabilities of proliferation,clone formation of MDA-MB-468 cells in transfection group decreased significantly compared with the non-transfection group and mock-vehicle group(P<0.05).Conclusion:CNTN1 gene is highly expressed in breast cancer MDA-MB-468 cells,and its silencing expression may inhibit proliferation and clone formation.

关 键 词：乳腺癌 MDA-MB-468 CNTN1 增殖 克隆形成 

分 类 号：R737.9[医药卫生—肿瘤]