摘要
目的探讨IL-6/JAK/STAT3和Smad3/TGF-β1信号通路在肝癌细胞上皮-间质转化(EMT)发生过程中的作用。方法分别予TGF-β1、IL-6、AG490刺激人肝癌细胞株(HepG2、Bel7402、MHCC97H、HCCLM3)后,qPCR检测Snail、STAT3的mRNA表达水平,蛋白质免疫检测p-STAT3/STAT3、p-Smad3/Smad3、TGF-β1以及EMT相关分子标志物Snail、E-Cadherin、Vimentin的蛋白表达水平。结果 TGF-β1可以诱导HepG2细胞EMT的发生,下调E-Cadherin的表达,上调Vimentin、Snai的表达。IL-6在肝癌细胞中通过刺激STAT3,激活Smad3/TGF-β1通路,进而上调EMT相关分子标志物Snail、Vimentin的表达。AG490(一种JAK2的特异性抑制剂)抑制p-STAT3/STAT3、p-Smad3及EMT相关分子标志物Snail的表达。结论STAT3在TGF-β1诱导的肝癌细胞EMT发生过程中作为一个正向调控因子,IL6/JAK/STAT3和Snail/Smad3/TGF-β1信号通路协同促进肝癌细胞EMT的发生。
Objective To investigate the underlying molecular mechanisms of interleukin-6/Janus kinase/signal transducer and activator of transcription(IL-6/JAK/STAT3)and Smad3/transforming growth factor-β1(Smad3/TGF-β1)signaling pathways during the epithelial to mesenchymal transition(EMT)process in hepatocellular carcinoma(HCC)cells.Methods Human HCC cell lines HepG2 and HCCLM3 were treated with TGF-β1,AG490 and IL-6,respectively.mRNA expressions of STAT3 and Snail were measured using quantitative PCR.Expressions of p-STAT3/STAT3,p-Smad3/Smad3,TGF-β1 and EMT-related markers were measured using western blotting.Results TGF-β1 induced EMT in HepG2 cells,dampen E-Cadherin expression and up-regulate expressions of vimentin,Snail,p-Smad2/3 and p-STAT3/STAT3.IL-6 activated Smad3/TGF-β1 pathway by stimulating STAT3 in human HCC lines,and up-regulated expressions of EMT-related molecular markers(Snail and Vimentin).AG490,the JAK2-specific inhibitor,inhibited expressions of pSTAT3/STAT3,p-Smad3 and Snail.Conclusion STAT3 aggravated TGF-β1-induced EMT and HCC metastasis,which performed a synergistical effect with IL-6/JAK/STAT3 and Snail/Smad3/TGF-β1 signaling pathways.
出处
《肝脏》
2018年第2期128-132,共5页
Chinese Hepatology
基金
国家自然基金面上项目(81570547)
国家科技部十二五项目(2012ZX10002007-001-040
2013ZX10002004-002-003)
院优秀青年培养计划(061405)
关键词
肝细胞癌
上皮-间质转化
信号转导和转录活化蛋白3
转化生长因子-Β1
Hepatocellular carcinoma
Epithelial-to-mesenchymal transition
Signal transducer and activator of transcription 3
Transforming growth factor-β1
