miR-141的过表达抑制MHCC-97H肝癌细胞的增殖和侵袭迁移  

Over-expression of miR-141 inhibits the proliferation,invasion and migration of hepatocellular carcinoma MHCC-97H cells

作　　者：姚博文[1] 薛雨墨 刘志奎[1] 徐蒙[1] 涂康生[1] 王军[2] 

机构地区：[1]西安交通大学第一附属医院肝胆外科,陕西西安710061 [2]西安交通大学第一附属医院急诊科,陕西西安710061

出　　处：《细胞与分子免疫学杂志》2016年第8期1083-1087,1093,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81402039)

摘　　要：目的观察肝细胞癌组织中miR-141的表达水平,通过上调MHCC-97H肝癌细胞中miR-141的水平观察对MHCC-97H细胞增殖、侵袭及迁移的影响。方法实时定量PCR检测miR-141在肝癌及癌旁组织中的表达水平,并分析miR-141表达水平与临床病理指标的关系及生存率。利用人工合成的miR-141模拟物,瞬时转染MHCC-97H细胞。MTT法检测MHCC-97H细胞的增殖,TranswellTM实验检测MHCC-97H细胞的侵袭及迁移,Westernblot法及免疫组织化学检测miR-141下游潜在靶点肝细胞产生的促红素受体A2(EphA2)的表达。结果miR-141在肝癌组织中表达水平显著低于相应癌旁组织;肝癌组织中miR-141低表达与TNM分期、门静脉侵犯及Edmondson分级显著相关;低表达miR-141患者3年生存率明显低于高表达组;miR-141模拟物转染可上调MHCC-97H细胞miR-141水平,上调miR-141可显著抑制MHCC-97H细胞的增殖、侵袭及迁移,并下调EphA2的蛋白水平;miR-141低表达组EphA2蛋白水平明显高于miR-141高表达组,相关性分析显示肝癌组织中miR-141与EphA2蛋白表达呈显著负相关。结论miR-141在肝癌组织中表达下调且与EphA2表达呈显著负相关,其低表达与肝癌恶性临床病理特征有关,上调miR-141表达可抑制EphA2的表达抑制肝癌细胞增殖、侵袭及迁移能力。Objective TO observe the expression level of miR-141 in tumor tissues of human hepatocellular carcinoma （HCC） and determine the effect of miR-141 level on cell proliferation, invasion and migration of MHCC-97H cells by upregulation of miR-141. Methods We checked the miR-141 expression level in HCC by real-time quantitative PCR and analyzed the relationship between the expression level of miR-141 and clinical pathological indicators as well as survival rate. MHCC-97H cells were transiently transfected with miR-141 mimics which were artificially synthesized. The proliferation of MHCC-97H cells was detected by MTT assay. TranswellTM assay was performed to examine the invasion and migration of MHCC-97H cells. The expression of erythropoietin-producing hepatocellular receptor A2 （ EphA2）, which was the potential downstream target, was determined by Western blotting and immunohistochemistry. Results The expression level of miR-141 in HCC tissues was significantly lower than that in the adjacent normal tissues, and it was obviously associated with TNM stage, portal vein infiltration and Edmondson degree. Patients in the lower miR-141 group had a worse 3-year survival than those in higher miR-141group. Overexpression of miR-141 in MHCC-97H cells significantly suppressed cell proliferation, invasion and migration, and inhibited the protein expression of EphA2. Correlation analysis showed that miR-141 level was negatively correlated with EphA2 expression level. Conclusion miR-141 is down-regulated in HCC tissues and it is negatively correlated with EphA2 expression. Its low expression is correlated with the malignant clinical pathological features, miR-141 overexpression down-regulates EphA2 expression and subsequently inhibits the proliferation, invasion and migration of HCC cells.

关 键 词：miR-141 肝细胞癌 增殖 侵袭 迁移 肝细胞产生的促红素受体A2(EphA2) 

分 类 号：R735.7[医药卫生—肿瘤]