miR-191靶向调控SATB1对肝癌细胞增殖和侵袭的影响  

Effects of miR-191 regulating SATB1 expression on proliferation and invasion of hepatoma cells

作　　者：陈红英 高贵峰 闫少茹[2] 王春艳 王长友 CHEN Hongying;GAO Guifeng;YAN Shaoru;WANG Chunyan;WANG Changyou(Department of General Surgery,Tangshan Hospital of Traditional Chinese Medicine,Tangshan 060300,China;不详)

机构地区：[1]唐山市中医医院普外科,河北唐山060300 [2]唐山市人民医院骨科 [3]华北理工大学附属医院普外科

出　　处：《山东医药》2022年第1期39-43,共5页Shandong Medical Journal

基　　金：河北省医学科学研究重点课题计划(201710096)。

摘　　要：目的探讨微小RNA-191(miR-191)靶向调控特异AT序列结合蛋白1(SATB1)对肝癌细胞增殖和侵袭的影响。方法体外传代培养人肝癌细胞系HepG2、人肝正常细胞系L02(以下分别称HepG2、L02细胞)。采用RT-PCR技术检测两种细胞miR-191、SATB1 mRNA表达,采用Western blotting法检测两种细胞SATB1蛋白表达。借助在线生物信息学软件starBase和TargetScan7.2预测miR-191与SATB1的结合位点,通过双荧光素酶报告基因实验验证miR-191与SATB1的靶向调控关系。取HepG2细胞,随机分为对照组、SATB1组、miR-191 mimic组、SATB1+miR-191 mimic组,对照组转染阴性对照质粒,SATB1组转染SATB1过表达质粒,miR-191 mimic组转染miR-191 mimic,SATB1+miR-191 mimic组转染SATB1过表达质粒和miR-191 mimic。收集各组细胞,采用MTT法检测细胞增殖能力,采用Transwell侵袭实验检测细胞侵袭能力,采用Western blotting法检测增殖相关蛋白Cyclin D1、p27及上皮间质转化相关蛋白E-cadherin、N-cadherin、Vimentin表达。结果HepG2细胞miR-191相对表达量低于L02细胞,而SATB1 mRNA和蛋白相对表达量均高于L02细胞(P均<0.01)。经在线生物信息学软件starBase和TargetScan7.2预测和双荧光素酶报告基因实验验证,miR-191与SATB1存在靶向调控关系。培养72 h,miR-191 mimic组细胞增殖、侵袭能力均低于对照组和SATB1组(P均<0.05);而SATB1+miR-191 mimic组细胞增殖、侵袭能力均高于miR-191 mimic组(P均<0.05)。与对照组和SATB1组比较,miR-191 mimic组Vimentin、N-cadherin、Cyclin D1蛋白相对表达量明显降低,E-cadherin、p27蛋白相对表达量明显升高(P均<0.05);与miR-191 mimic组比较,SATB1+miR-191 mimic组Vimentin、N-cadherin、Cyclin D1蛋白相对表达量明显升高,E-cadherin、p27蛋白相对表达量明显降低(P均<0.05)。结论miR-191通过靶向负调控SATB1抑制肝癌细胞增殖和侵袭,其机制可能与调控增殖相关蛋白Cyclin D1、p27及上皮间质转化相关蛋白E-cadherin、N-cadherin、VimObjective To explore the effects of microRNA-191(miR-191)regulating specific AT sequence binding protein 1(SATB1)on the proliferation and invasion of hepatoma cells.Methods Human hepatoma cell line HepG2 and human normal liver cell line L02(hereinafter referred to as HepG2 and L02 cells,respectively)were cultured in vitro.The expression levels of miR-191 and SATB1 mRNA were detected by RT-PCR,and the expression of SATB1 protein was detected by Western blotting.The interaction sites between miR-191 and SATB1 were predicted by online bioinformatics software starBase and TargetScan 7.2,and the targeted regulation relationship between miR-191 and SATB1 was verified by double luciferase reporter gene experiment.HepG2 cells were taken and randomly divided into the control group,SATB1 group,miR-191 mimic group,and SATB1+miR-191 mimic group.The cells in the control group were transfected with negative control plasmid,the SATB1 group with SATB1 overexpression plasmid,the miR-191 mimic group with miR-191 mimic,and the SATB1+miR-191 mimic group with SATB1 overexpression plasmid and miR-191 mimic,respectively.The cells of each group were collected;the cell proliferation ability was detected by MTT,the cell invasion ability was detected by Transwell invasion test,and the expression levels of Cyclin D1 protein(CD1P),p27 protein,epithelial cadherin(E-cad),neural cadherin(N-cad)and vimentin(Vim)proteins were detected by Western blotting.Results The relative expression of miR-191 in HepG2 cells was lower than that in L02 cells,while the relative expression levels of SATB1 mRNA and protein were higher than those in L02 cells(all P<0.01).Through the prediction of online bioinformatics software starBase and Targetscan 7.2 prediction and the verification of double luciferase reporter gene experiment,miR-191 had a targeted regulatory relationship with SATB1.After 72 h of culture,the cell proliferation and invasion abilities of the miR-191 mimic group were lower than those of the control group and SATB1 group(all P<0.05),while the cell

关 键 词：肝癌 微小RNA-191 特异AT序列结合蛋白1 细胞增殖 细胞侵袭 

分 类 号：R735.7[医药卫生—肿瘤]