miR-147a在宫颈癌Hela细胞中的表达及对Hela 细胞增殖、黏附及迁移的调控作用  

Expression of miR-147a in cervical cancer Hela cells and its regulation role on proliferation,adhesion and migration of Hela cells

作　　者：郭健 曾友玲 董晓寒 GUO Jian;ZENG Youling;DONG Xiaohan(Department of Gynecology and Obstetrics,Wuhan Children′s Hospital(Wuhan Municipal Maternal and Child Healthcare Hospital),Tongji Medical College,Huazhong University of Science&Technology,Wuhan,Hubei 430022,China)

机构地区：[1]华中科技大学同济医学院附属武汉儿童医院(武汉市妇幼保健院)妇产科,湖北武汉430022

出　　处：《检验医学与临床》2024年第14期2047-2052,共6页Laboratory Medicine and Clinic

基　　金：湖北省武汉市卫生和计划生育委员会科研项目(WX17D16)。

摘　　要：目的探讨微小核糖核酸-147a(miR-147a)在宫颈癌Hela细胞中的表达及对宫颈癌Hela细胞增殖、黏附及迁移的调控作用。方法在体外培养人宫颈癌Hela细胞,将对数期生长细胞分为对照组(不进行干预)、mimics NC组(转染mimics NC片段至Hela细胞)、inhibitor NC组(转染inhibitor NC片段至Hela细胞)、miR-147a mimics组(转染miR-147a mimics片段至Hela细胞)及miR-147a inhibitor组(转染miR-147a inhibitor片段至Hela细胞),干预24 h。采用实时荧光定量聚合酶链反应检测Hela细胞中miR-147a表达水平;采用细胞计数试剂盒-8测定细胞活力;采用5-乙炔基-2′脱氧尿嘧啶核苷测定细胞的增殖率;采用细胞黏附实验测定细胞的黏附能力;采用Transwell小室法测定细胞的迁移能力;采用蛋白免疫印迹法测定细胞周期蛋白D1(cyclin D1)和核转录因子-κB(NF-κB)通路相关蛋白表达水平。结果miR-147a mimics组miR-147a表达水平明显高于mimics NC组(P<0.05),miR-147a inhibitor组miR-147a表达水平明显低于inhibitor NC组(P<0.05),miR-147a mimics、miR-147a inhibitor转染成功。与mimics NC组相比,miR-147a mimics组细胞活力、增殖率、黏附数、迁移数及cyclin D1、磷酸化(p)-NF-κB蛋白表达水平明显下降(P<0.05);与inhibitor NC组相比,miR-147a inhibitor组细胞活力、增殖率、黏附数、迁移数及cyclin D1、p-NF-κB蛋白表达水平明显升高(P<0.05)。结论过表达miR-147a通过调控NF-κB通路抑制宫颈癌细胞的增殖、黏附及迁移能力。Objective To investigate the expression of microRNA-147a(miR-147a)in cervical cancer Hela cells and its regulatory effect on the proliferation,adhesion and migration of cervical cancer Hela cells.Methods The human cervical cancer Hela cells were cultured in vitro.The logarithmic growth cells were divided into the control group(without intervention),mimics NC group(transfecting mimics NC fragments into Hela cells),inhibitor NC group(transfecting inhibitor NC fragments into Hela cells)and miR-147a mimics group(transfecting miR-147a mimics fragments into Hela cells)and miR-147a inhibitor group(transfected miR-147a inhibitor fragments into Hela cells).The intervention lasted for 24 h.The real-time fluorescence quantitative PCR was used to determine the expression level of miR-147a in Hela cells.The cell counting Kit-8 was used to measure the cell viability.The cell proliferation rate was determined by 5-acetyne-2′deoxyuracil nucleoside.The cell adhesion ability was determined by cell adhesion assay.The Transwell chamber method was used to measure the migration ability of cells.The expression levels of Cell cycle protein D1(cyclinD 1)and NF-κB pathway related proteins were determined by Western blot.Results The expression level of miR-147a in the miR-147a mimics group was significantly higher than that in the mimics NC group(P<0.05),the expression level of miR-147a in the miR-147a inhibitor group was significantly lower than that in the inhibitor NC group(P<0.05).The transfection of miR-147a mimics and miR-147a inhibitor was successful.Compared with the mimics NC group,the cell viability,proliferation rate,adhesion number,migration number and the expression levels of cyclin D1 and phosphorylated(p)-NF-κB protein in the miR-147a mimics group were decreased significantly(P<0.05).Compared with the inhibitor NC group,the cell viability,proliferation rate,adhesion number,migration number and cyclin D1 and p-NF-κB protein expression levels of the miR-147a inhibitor group were significantly increased(P<0.05).Conclusion

关 键 词：宫颈癌 微小核糖核酸-147a 核转录因子-κB信号通路 增殖 黏附 迁移 

分 类 号：R737.33[医药卫生—肿瘤] R446.6[医药卫生—临床医学]