miRNA-129-5p干扰技术对胆道闭锁小鼠胆道EMT作用机制的研究
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摘要
目的探讨miRNA-129-5p在胆道闭锁上皮间质转化(EMT)中的作用机制。方法构建胆道闭锁小鼠模型(实验组),取实验组胆道组织,构建TGF-β1诱导的胆管上皮EMT细胞模型,检测TGF-β1诱导刺激后胆道上皮细胞株EMT相关标志物表达水平,并检测miRNA-129-5p的表达变化。同时转染miRNA-129-5p的前体,比较转染前后TGF-β1刺激下胆道上皮细胞EMT相关标志物及细胞外基质的表达改变。结果实验组小鼠肝脏肝外胆道呈EMT样改变,miRNA-129-5p在实验组小鼠中的表达水平下调(P<0.05),过表达miRNA-129-5p可以抑制EMT的进程(P<0.05)。结论 miRNA-129-5p可能通过TGF-β1通路调控胆道上皮EMT的进程。
Objective To investigate the mechanism of miRNA-129-5 p in epithelial-mesenchymal transition(EMT)of biliary atresia.Methods Constructed bile duct epithelial EMT cell model(the experimental group)induced by TGF-β1,detected the expressions of EMT related markers and miRNA-129-5 p.While miRNA-129-5 p precursor was transfected,the expressions of EMT related markers and extracellular matrix were contrasted between the original and the renovated biliary epithelial cells.Results In the experimental group,extrahepatic bile duct showed EMT,the expression of miRNA-129-5 p was decreased(P<0.05),overexpression of miRNA-129-5 p could inhibit the progression of EMT(P<0.05).Conclusion miRNA-129-5 p may relate to EMT by regulating the expression of TGF-β1.
Objective To investigate the mechanism of miRNA-129-5 p in epithelial-mesenchymal transition(EMT)of biliary atresia.Methods Constructed bile duct epithelial EMT cell model(the experimental group)induced by TGF-β1,detected the expressions of EMT related markers and miRNA-129-5 p.While miRNA-129-5 p precursor was transfected,the expressions of EMT related markers and extracellular matrix were contrasted between the original and the renovated biliary epithelial cells.Results In the experimental group,extrahepatic bile duct showed EMT,the expression of miRNA-129-5 p was decreased(P<0.05),overexpression of miRNA-129-5 p could inhibit the progression of EMT(P<0.05).Conclusion miRNA-129-5 p may relate to EMT by regulating the expression of TGF-β1.
引文
[1]HARADA K.Sclerosing and obstructive cholangiopathy in biliary atresia:mechanisms and association with biliary innate immunity[J].Pediatr Surg Int,2017,33(12):1243-1248.
[2]HSU Y A,LIN C H,LIN H J,et al.Effect of microRNA-155on the interferon-gamma signaling pathway in biliary atresia[J].Chin J Physiol,2016,59(6):315-322.
[3]YE Y,LI Z,FENG Q,et al.Downregulation of microRNA-145may contribute to liver fibrosis in biliary atresia by targeting ADD3[J].PLoS One,2017,12(9):e0180896.
[4]GOLDMAN M,PRANIKOFF T.Biliary disease in children[J].Curr Gastroenterol Rep,2011,13(2):193-201.
[5]MA N,CHEN F,SHEN S L,et al.MiRNA-129-5p inhibits hepatocellular carcinoma cell metastasis and invasion via targeting ETS1[J].Biochem Biophys Res Commun2015,461(4):618-623.
[6]ZHANG J,LI S,YAN Q,et al.Interferon-βinduced miRNA-129-5p down-regulates HPV-18E6and E7viral gene expression by targeting SP1in cervical cancer cells[J].PLoS One,2013,8(12):e81366.
[7]DONG R,SHEN Z,ZHENG C,et al.Serum microRNA microarray analysis identifies miR-4429and miR-4689are potential diagnostic biomarkers for biliary atresia[J].Sci Rep,2016,16(6):21084.
[8]GOLDSCHMIDT I,THUM T,BAUMANN U.Circulating miR-21and miR-29aas markers of disease severity and etiology in cholestatic pediatric liver disease[J].J Clin Med,2016,5(3):e28.
[9]SHAN Y,SHEN N,HAN L,et al.MicroRNA-499Rs3746444polymorphism and biliary atresia[J].Dig Liver Dis,2016,48(4):423-428.
[10]杨瑛,汤绍涛,曹国庆,等.L-12P35,P40和IFN-γ在胆道闭锁小鼠模型肝脏组织中的表达[J].临床小儿外科杂志,2008,7(6):26-29.
[2]HSU Y A,LIN C H,LIN H J,et al.Effect of microRNA-155on the interferon-gamma signaling pathway in biliary atresia[J].Chin J Physiol,2016,59(6):315-322.
[3]YE Y,LI Z,FENG Q,et al.Downregulation of microRNA-145may contribute to liver fibrosis in biliary atresia by targeting ADD3[J].PLoS One,2017,12(9):e0180896.
[4]GOLDMAN M,PRANIKOFF T.Biliary disease in children[J].Curr Gastroenterol Rep,2011,13(2):193-201.
[5]MA N,CHEN F,SHEN S L,et al.MiRNA-129-5p inhibits hepatocellular carcinoma cell metastasis and invasion via targeting ETS1[J].Biochem Biophys Res Commun2015,461(4):618-623.
[6]ZHANG J,LI S,YAN Q,et al.Interferon-βinduced miRNA-129-5p down-regulates HPV-18E6and E7viral gene expression by targeting SP1in cervical cancer cells[J].PLoS One,2013,8(12):e81366.
[7]DONG R,SHEN Z,ZHENG C,et al.Serum microRNA microarray analysis identifies miR-4429and miR-4689are potential diagnostic biomarkers for biliary atresia[J].Sci Rep,2016,16(6):21084.
[8]GOLDSCHMIDT I,THUM T,BAUMANN U.Circulating miR-21and miR-29aas markers of disease severity and etiology in cholestatic pediatric liver disease[J].J Clin Med,2016,5(3):e28.
[9]SHAN Y,SHEN N,HAN L,et al.MicroRNA-499Rs3746444polymorphism and biliary atresia[J].Dig Liver Dis,2016,48(4):423-428.
[10]杨瑛,汤绍涛,曹国庆,等.L-12P35,P40和IFN-γ在胆道闭锁小鼠模型肝脏组织中的表达[J].临床小儿外科杂志,2008,7(6):26-29.