miR-21在TGF-β1引起的肝卵圆细胞上皮间质转化中的作用  

Effects of miR-21 on TGF-β1 induced epithelial-mesenchymal transition of hepatic oval cells

作　　者：高胜强 喻凯[1] 陈潇[1] 罗建生[1] GAO Sheng-qiang;YU Kai;CHEN Xiao;LUO Jian-sheng(Department of Hepatobiliary and Pancreatic Surgery,Jinhua Municipal Hospital,Jinhua,Zhejiang 321000,China)

机构地区：[1]金华市中心医院肝胆胰外科,浙江金华321000

出　　处：《肝胆胰外科杂志》2018年第2期142-147,共6页Journal of Hepatopancreatobiliary Surgery

基　　金：金华市科技局项目(2017-4-007)

摘　　要：目的探讨microRNA-21(miR-21)在TGF-β1引起的肝卵圆细胞(hepatic oval cell,HOC)上皮间质转化(epithelial mesenchymal transition,EMT)中的作用。方法体外培养大鼠肝卵圆细胞,用10 ng/mL TGF-β1刺激HOC 1、3、5、7 d后,Western blotting检测EMT相关蛋白(E-cadherin、N-cadherin和Vimentin)表达情况;用10 ng/mL TGF-β1刺激HOC 5 d后,实时荧光定量PCR检测miR-21的表达情况;分别用miR-21增强和抑制慢病毒转染HOC构建稳定的细胞株,PCR检测miR-21的表达情况;接着用TGF-β1刺激miR-21抑制慢病毒转染的HOC 5 d,Western blotting检测E-cadherin、N-cadherin和Vimentin的表达;用miR-21增强慢病毒转染HOC后,Western blotting检测E-cadherin、N-cadherin和Vimentin的表达。结果 TGF-β1刺激HOC 1、3、5、7 d后,E-cadherin逐渐减低,N-cadherin和Vimentin逐渐升高。TGF-β1刺激HOC 5 d后miR-21的表达增强了4.32倍。分别用增强和抑制慢病毒转染HOC后,miR-21的表达增强了3.82倍和抑制了0.22倍。下调miR-21表达后,TGF-β1致HOC的EMT作用减弱。miR-21过表达后,HOC发生了EMT。结论抑制miR-21的表达可以减少TGF-β1引起的HOC的EMT作用,从而减轻肝纤维化。Objective To investigate the role of microRNA-21(miR-21)in the epithelial mesenchymal transition(EMT)of hepatic oval cells(HOC)induced by TGF-β1.Methods Hepatic oval cells were cultured in vitro.The EMT-related proteins including E-cadherin,N-cadherin and Vimentin were detected by Western blotting after stimulation with 10 ng/mL TGF-β1 for 1,3,5 and 7 d.The expression of miR-21 was detected by real-time quantitative PCR with 10 ng/mL TGF-β1 for 5 d.The stable cell lines were constructed with enhanced and inhibited lentivirus transfection respectively and the expression of miR-21 was detected by PCR.The expression of E-cadherin,N-cadherin and Vimentin was detected by Western blotting after TGF-β1 stimulation on hepatic oval cells with miR-21 inhibited lentivirus transfection for 5 d.The expression of E-cadherin,N-cadherin and Vimentin was detected by Western blotting on hepatic oval cells with miR-21 enhanced lentivirus transfection.Results After the hepatic oval cells were stimulated by TGF-β1 for 1,3,5 and 7 d,the expression of E-cadherin decreased gradually,accompanying with gradually increased expression of N-cadherin and Vimentin.The expression of miR-21 in hepatic oval cells increased by 4.32 times after TGF-β1 stimulation for 5 d.The expression of miR-21 was enhanced by 3.82 times and decreased by 0.22 times respectively after the enhancement and inhibition of lentivirus transfection on hepatic oval cells.The down-regulated expression of miR-21 alleviated the EMT effect induced by TGF-β1on hepatic oval cells.After over-expression of miR-21,epithelial mesenchymal transition occurred in hepatic oval cells.Conclusion The inhibited expression of miR-21 can reduce the EMT effect induced by TGF-β1stimulation on hoc,thereby reducing liver fibrosis.

关 键 词：MICRORNA-21 肝卵圆细胞 转化生长因子Β1 上皮-间质转化 

分 类 号：R575[医药卫生—消化系统]