miRNA-628-3p调控乳腺癌细胞增殖的作用及机制研究  

Effect and mechanism of miR-628-3p on proliferation of breast cancer cells

作　　者：董文珠 陈杭萍 陈赛贞[1] 夏哲林 章欣[1] 姚立[2] DONG Wenzhu;CHEN Hangping;CHEN Saizhen;XIA Zhelin;ZHANG Xin;YAO Li(Department of Pharmacy,Taizhou Central Hospital,Taizhou,Zhejiang 318000,China;Institute of Pharmacy,College of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou,Zhejiang 310053,China)

机构地区：[1]台州市中心医院药剂科,浙江台州318000 [2]浙江中医药大学药学院研究所,杭州310053

出　　处：《重庆医学》2019年第18期3082-3086,共5页Chongqing medicine

基　　金：浙江中医药大学校级科研基金项目(2016ZG11)

摘　　要：目的探讨miRNA-628-3p对乳腺癌细胞增殖的影响及其可能的机制。方法以反转录PCR(RT-PCR)方法检测人正常乳腺上皮细胞(MCF-10A)与乳腺癌细胞(MCF-7、MDA-MB-231)中miR-628-3p的表达水平,检测miRNA-628-3p在MCF-7细胞中的转染效率;噻唑蓝(MTT)法检测miRNA-628-3p对MCF-7细胞增殖的影响;生物信息学对miRNA-628-3p进行靶基因预测,并通过荧光素酶报告基因实验验证,Westernblot检测过表达miRNA-628-3p对靶基因多聚腺苷酸结合蛋白互作蛋白1(PAIP1)蛋白表达水平的影响;MTT法检测抑制靶基因PAIP1表达后对MCF-7细胞增殖的影响。结果乳腺癌细胞中miRNA-628-3p的表达水平明显低于人正常乳腺上皮细胞;与空载体阴性对照组(NC)比较,miRNA-628-3p过表达组细胞增殖能力明显降低;生物信息学分析表明PAIP1是miRNA-628-3p直接作用的靶基因,荧光素酶报告基因验证了该结果,并且过表达miRNA-628-3p可以明显下调PAIP1的蛋白表达水平;抑制靶基因PAIP1的表达能抑制MCF-7细胞的增殖。结论miRNA-628-3p在乳腺癌细胞中低表达,可以通过靶向调控PAIP1的表达进而抑制乳腺癌细胞的增殖。Objective To investigate the effect of miR-628-3p on the proliferation of breast cancer cells and its possible mechanism. Methods The expressions of miR-628-3p in human normal breast epithelial cells (MCF-10A) and breast cancer cells (MCF-7 and MDA-MB-231) were detected by RT-PCR,and the transfection efficiency of miR-628-3p in MCF-7 cells was detected.MTT assay was used to detect the effect of miR-628-3p on the proliferation of MCF-7 cells.The target gene of miR-628-3p was predicted by bioinformatics,and verifies by luciferase reporter gene experiment.The effect of miR-628-3p overexpression on the protein expression of target gene Poly(A)-binding protein-interacting protein 1 (PAIP1) was detected by Western blot.MTT assay was used to detect the proliferation of MCF-7 cells after inhibiting the expression of target gene PAIP1. Results The expression of miR-628-3p in breast cancer cells was significantly lower than that in human normal breast epithelial cells.Compared with the empty carrier negative control group (NC),the cell proliferation was significantly reduced in the miR-628-3p overexpression group.Bioinformatics analysis showed that PAIP1 was a direct target gene of miR-628-3p,which was verified by luciferase reporter gene experiment.And the overexpression of miR-628-3p could significantly down-regulate the expression of PAIP1 protein.Inhibiting the expression of the target gene PAIP1 also inhibited the proliferation of MCF-7 cells. Conclusion miR- 628-3p is low-expressed in breast cancer cells and can inhibit the proliferation of breast cancer cells by target regulating the expression of PAIP1.

关 键 词：miRNA-628-3p 乳腺癌细胞 细胞增殖 机制 

分 类 号：R966[医药卫生—药理学]