MRPS23 shRNA对乳腺癌细胞Walker256增殖凋亡的作用及机制  

Effects and Mechanisms of MRPS23-targeted shRNA on Proliferation and Apoptosis of Breast Cancer Cell Walker256

作　　者：高燕[1] 李伏燕[1] 李薇[1] 陈义加[1] 吴瑞敏[1] 周红[1] 杨怡[1] 裴之俊[1] 

机构地区：[1]十堰市太和医院.湖北医药学院附属医院PET中心,湖北十堰442000

出　　处：《湖北医药学院学报》2016年第6期527-530,546,共5页Journal of Hubei University of Medicine

基　　金：国家自然科学基金青年项目(81401447);十堰市科技局指导项目(2016Y16);湖北医药学院重点专科建设项目

摘　　要：目的:探讨线粒体核糖体蛋白MRPS23 shRNA对大鼠乳腺癌细胞Walker256增殖和凋亡的作用及机制。方法:构建含有MRPS23基因沉默片段(sh MRPS23)和对照shRNA的慢病毒载体(sh Ctrl),感染Walker256细胞48 h,以确定转染效率和基因沉默效率。MTS、TUNEL法检测其对Walker256细胞的增殖凋亡率的影响;RTPCR、western blot法检测细胞增殖和凋亡相关基因与蛋白表达水平。结果:与control组以及对照病毒组相比,sh MRPS23能显著抑制MRPS23基因及蛋白的表达,MRPS23基因沉默后能抑制Walker256细胞增殖,并促进细胞凋亡;p21WAF1、p53表达增高(P<0.05)。结论:慢病毒介导的MRPS23 shRNA可以通过上调p21WAF1抑制Walker256细胞增殖,MRPS23 RNAi可诱导细胞p53依赖性的凋亡。Objective To investigate the effects and mechanisms of mitoehondrial ribosomal proteins （MRPS23）-targeted shRNA on the proliferation and apoptosis of breast cancer cells Walker256. Methods The shRNA targeting against MRPS23 （shMRPS23） or non-silence sequence （shCtrl） Was suhcloned into lentivirus vector that containing green fluorescent protein （GFP） DNA sequence. Rat breast cancer cells Walker256 were respectively transfected with lentivirus （shCtrl, shMRPS23） or PBS （Control） for 48 h, so as to confirm the efficiency of transfeetion and gene silencing. The proliferation of cells was analyzed by MTT assay. The cell apoptosis was quantified TUNEL staining. The expression of MRPS23 and apoptosis-associated genes in Walker256 were detected by RT-PCR and Western blot in different groups： Results Compared with controls, the expression of MRPS23 was significantly inhibited by shMRPS23 in Walker256 cells. MRPS23 shRNA could inhibit the proliferation of breast cancer cell and significantly increased apoptotic rate （P〈0.05）. QPCR and western blot demonstrated that shMRPS23 could increase the expression of p53 and p21 WAF1/CIP1. Conclusion Lentivirus mediated MRPS23 shRNA can inhibit the proliferation of breast cancer cells, which may be related with the activation of p21 WAFI/CIPI. Additionally, the p53-dependent apoptotic pathway was induced by shMRPS23.

关 键 词：MRPS23 SHRNA 乳腺癌细胞 细胞增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]