摘要
目的探究苦参素(OM)对转化生长因子-β1(TGF-β1)诱导人肝癌细胞HepG2上皮细胞间充质转化(EMT)过程的影响及其分子机制。方法用10 ng/ml TGF-β1诱导HepG2细胞建立EMT模型,将建模成功的肝癌细胞分为5组:模型组、OM 1 mg/ml组、OM 2 mg/ml组、OM 4 mg/ml组、OM 8 mg/ml组,以未建模的HepG2细胞(空白组)作为对照组,采用实时荧光定量PCR(qRT-PCR)检测HepG2中miR-204表达情况,分别采用CCK8实验、划痕实验检测HepG2细胞增殖、迁移情况,蛋白免疫印迹(Western Blot)检测HepG2细胞中钙黏附蛋白-E(E-cadherin)、波形蛋白(Vimentin)表达情况。结果模型组HepG2细胞呈现梭形和纺锤形,且细胞间隙变大、排列松散,表明造模成功。与空白组比较,模型组HepG2细胞中miR-204表达水平、HepG2细胞生长抑制率、迁移能力及Vimentin表达水平高于空白组(P<0.05);与模型组相比,OM 4 mg/ml组HepG2细胞中miR-204表达水平、HepG2细胞增殖率、迁移能力及Vimentin表达水平降低,E-cadherin表达水平升高(P<0.05)。结论OM可上调HepG2细胞中miR-204表达,进而抑制TGF-β1诱导的人肝癌细胞HepG2 EMT过程。
Objective To investigate the effect of oxymatrine(OM)on epithelial-mesenchymal transition(EMT)induced by the transforming growth factor-β1(TGF-β1)in human hepatoma cell line HepG2 and related molecular mechanism.Methods 10 ng/ml TGF-β1 was used to induce HepG2 cells to establish EMT model.The hepatoma carcinoma cells(HCC)successfully modeled were divided into 5 groups:the model group,1 mg/ml OM group,2 mg/ml OM group,4 mg/ml OM group,and 8 mg/ml OM group.Unmodeled HepG2 cells(blank group)were used as the control group.The expression of miR-204 in HepG2 was detected by real-time fluorescence quantitative PCR(qRT-PCR).The proliferation and migration of HepG2 cells were detected by CCK8 experiment and scratch experiment,respectively.The expressions of E-cadherin and Vimentin in HepG2 cells were detected by Western Blot experiment.Results The HepG2 cells in the model group were characterized by fusiform and spindle shapes,and their cell gap was enlarged and loosely arranged,indicating the success of modeling.Compared with those of the blank group,the miR-204 expression,growth inhibition rate and migration ability of HepG2 cells and Vimentin expressions in the model group were higher than those in the blank group(P<0.05).Compared with those of the model group,the miR-204 expression,proliferation rate and migration ability of HepG2 cells and Vimentin expressions in the 4 mg/ml OM group decreased,while the E-cadherin expression increased(P<0.05).Conclusion OM can up-regulate the expression of miR-204 in HepG2 cells,thereby inhibiting the EMT process of human hepatoma HepG2 cells induced by TGF-β1.
作者
张彩灵
覃小珊
黄赞松
Zhang Cailing;Qin Xiaoshan;Huang Zansong(Graduate School,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;The Affiliated Hospital of Youjiang Medical University for Nationalities,Guangxi Clinical Medicine Research Center for Hepatobiliary Diseases,Baise 533000,Guangxi,China)
出处
《右江民族医学院学报》
2021年第1期11-16,共6页
Journal of Youjiang Medical University for Nationalities
基金
广西自然科学基金项目(2014GXNSFAA118143)
广西科技基地和人才专项(广西肝胆疾病临床医学研究中心研究课题)(桂科AD17129025)
2017年度广西医药卫生自筹经费计划课题(Z20170224)
2020年右江民族医学院学术学位研究生创新计划项目(YXCXJH2020009)。
