王金利;张敏;
WANG Jinli;ZHANG Min;the People's Hospital of Huishan District;Department of Pharmacology,Guizhou Medical University;

• 1:无锡市惠山区人民医院
• 2:贵州医科大学

摘要(Abstract)：

目的:探讨antagomiR-103协同阿霉素(ADM)对TGF-β1诱导肝癌细胞(HepG2)发生上皮-间质细胞转化(EMT)及HepG2细胞存活率的影响。方法:HepG2细胞体外培养,设为control组、TGF-β1组、TGF-β1+ADM组、TGF-β1+microRNA inhibitor N.C组、TGF-β1+ADM+microRNA inhibitor N.C组、TGF-β1+antagomiR-103组、TGF-β1+ADM+antagomiR-103组及使用western-blot检测EMT标志蛋白N-Cadherin、E-Cadherin、vimentin的表达水平;设control组、TGF-β1组、antagomiR-103组、TGF-β1+antagomiR-103组,用不同浓度的ADM作用于各组细胞24 h,使用MTT法检测细胞存活率,并计算IC50。结果:与control组相比,antagomiR-103+ADM组E-Cadherin表达上调,N-Cadherin、Vimentin表达下调;细胞存活率随ADM浓度的增高而逐渐降低;在ADM浓度相同的条件下,不同药物预处理组之间进行比较,当ADM浓度为25 mg/L时,antagomiR-103+ADM组的细胞存活率明显低于其他组。结论:antagomiR-103联合ADM可抑制TGF-β1诱导发生EMT的HepG2细胞存活。
Objective:To investigate the effect of antagomir-103 synergized by ADM on epithelial mesenchymal transition(EMT) in Hep G2 induced by TGF-β1 and on the cell survival rate of and Hep G2 cells.Methods:Hep G2 cells were cultivated in DMEM with 10% fetal calf serum and divided into the control group,the TGF-β1 group,TGF-β1+ADM group,TGF-β1+microRNA inhibitor N.C group,TGF-β1+ADM+microRNA inhibitor N.C group,the TGF-β1+antagomiR-103 group,and the TGF-β1+ADM+antagomiR-103 group.Western-blot was adopted to detect the expression level of EMT,E-cadherin,vimentin and N-cadherin in all these groups.On the other hand,the Hep G2 cells were also divided into the control group,TGF-β1 group,antagomiR-103 group,TGF-β1+antagomiR-103.With different concentrations of ADM on each above group for 24 h,MTT method was used to detect cell survival rate,and IC 50 was calculated.Results:Compared with control group,the expression of E-cadherin was up-regulated in antagomiR-103+ADM group,while N-cadherin,vimentin expression was down regulated.The cell survival rate decreased with the increase of ADM concentration.The cell survival rate of antagomiR-103+ADM group was significantly lower than that of the other groups at the same ADM concentration and when the ADM concentration was 25 mg/L.Conclusion:antagomiR-103 combined with ADM can inhibit transformation of TGF-β1 induced EMT and growth of Hep G2 cell.

关键词(KeyWords)： 药物疗法,联合;肝肿瘤;细胞;antagomiR-103;阿霉素;上皮细胞-间充质细胞转换;TGFβ-1
drug therapy;combination;liver neoplasms;cells;antagomiR-103;adriamycin;epithelial-mesenchymal transitions;TGFβ-1

Abstract:

Keywords:

基金项目(Foundation): 贵阳市科技局基金项目[(2011103)17号]

作者(Author): 王金利;张敏;
WANG Jinli;ZHANG Min;the People's Hospital of Huishan District;Department of Pharmacology,Guizhou Medical University;

Email:

DOI: 10.19367/j.cnki.1000-2707.2016.12.013

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