MiR-130a靶向调控CYLD基因及其对肝癌细胞增殖的影响  

MicroRNA-130a targets CYLD to regulate cell proliferation in hepatocellular carcinoma

作　　者：倪芳[1] 赵华[1] 汪心怡[1] 陈卓[1] 汪思应[1] 

机构地区：[1]安徽医科大学基础医学院病理生理学教研室,安徽省合肥市230032

出　　处：《世界华人消化杂志》2014年第11期1504-1509,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.31300715;安徽省自然科学基金资助项目;No.1308085QH136~~

摘　　要：目的:检测microRNA-130a(miR-130a)在肝癌中的表达水平及其对CYLD基因表达的调控,并探讨miR-130a对肝癌细胞增殖的影响.方法:采用qRT-PCR检测miR-130a在原发性肝癌组织及对应的癌旁组织、肝癌细胞系和正常肝细胞中的表达差异,利用靶基因预测分析软件预测miR-130a潜在靶基因CYLD后,构建携带靶基因CYLD野生型及突变型3'UTR序列的双荧光素酶报告基因质粒,应用双荧光素酶报告基因实验体系进行验证;采用miR-130a抑制剂下调肝癌细胞系HepG2中miR-130a的表达,Western blot检测CYLD蛋白的表达变化;MTT法检测肝癌细胞增殖的改变.结果:MiR-130a在原发性肝癌组织中的表达明显高于对应的癌旁组织(P<0.05),在肝癌细胞系中的表达也显著高于正常肝细胞(P<0.05);双荧光素酶报告基因系统验证CYLD是miR-130a的直接靶基因;HepG2细胞中转染miR-130a抑制剂下调miR-130a表达后.Western blot检测结果显示CYLD蛋白表达水平显著上调;miR-130a抑制剂转染的HepG2细胞其增殖受到明显抑制.结论:MiR-130a在肝癌中表达上调,下调miR-130a后可以促进靶基因CYLD的表达,并可以抑制肝癌细胞的增殖活性,miR-130a有可能成为原发性肝癌治疗的新靶点.AIM： To detect the expression level of miR-130a in hepatocellular carcinoma and to analyze the effect of miR-130a on hepatocellular carcinoma cell proliferation.METHODS： MiR-130a expression levels in hepatocellular carcinoma and non-tumor pericarcinous tissues, as well as normal liver and hepatocellular carcinoma cell lines were determined by qRT-PCR. The CYLD 3＇-UTR as the target of miR-130a was predicted by bioinformatics analysis. The reporter activity was evaluated by the dual luciferase reporter assay. After HepG2 cells were transfected with a synthetic miR-130a inhibitor （anti-miR-3666） or a synthetic control miRNA （anti-miR-C）, the expression of CYLD protein in the transfected HepG2 cells was determined by Western blot, and cell proliferation was determined by MTT assay.RESULTS： MiR-130a expression was up-regulated in hepatocellular carcinoma compared to non-tumor pericarcinous tissues. The results of dual luciferase assays validate CYLD as a specific target gene of miR-130a. Inhibition of miR-130a resulted in the upregulation of CYLD protein expression in HepG2 cell line. MiR-130a knockdown significantly inhibited the cell proliferation of HepG2 cells.CONCLUSION： MiR-130a is overexpressed in hepatocellular carcinoma. Inhibition of miR-130a promotes the proliferation of hepatocellular carcinoma cells. MiR-130a may become a new potential therapeutic target for hepatocellular carcinoma.

关 键 词：原发性肝细胞癌 MiRNA-130a CYLD 增殖 

分 类 号：R735.7[医药卫生—肿瘤]