微小RNA-181b及其靶基因肝癌缺失基因-1在肝癌发生中的作用  

Role of microRNA181-b and its target gene oncogene deleted in liver cancer-1 in hepatocellular carcinoma pathogenesis

作　　者：李颖[1] 卢忠心[2] 孔晓宇[3] 魏礼清[2] 

机构地区：[1]华中科技大学同济医学院附属武汉市中心医院,武汉430014 [2]华中科技大学同济医学院检验科,武汉430014 [3]华中科技大学同济医学院肝胆外科,武汉430014

出　　处：《中华实验外科杂志》2013年第10期2070-2072,共3页Chinese Journal of Experimental Surgery

基　　金：武汉市卫生局临床医学科研项目（WX12C52）

摘　　要：目的 探讨微小RNA（miRNA,miR）-181b对肝癌缺失基因-1（DLC-1）的调控及其在肝癌发病中的作用.方法 通过实时荧光定量聚合酶链反应（FQ-PCR）和Western blot检测4份正常肝脏组织,17份癌旁组织、肝癌组织及肝癌SMMC7721细胞、胚肝LO2细胞中miR-181b和DLC-1的mRNA和蛋白质表达水平.应用miR-181b抑制剂下调肝癌细胞SMMC7721中miR-181b表达后,实时定量聚合酶链反应和Westem blot检测SMMC7721细胞中DLC-1表达水平的变化.结果 miR-181b在正常肝组织、癌旁组织和肝癌组织中的相对表达量分别为0.162 ±0.004、0.175±0.021和0.656±0.034,肝癌组织高于正常肝组织（P＜0.05）.miR-181b在SMMC7721细胞中的表达较LO2细胞上调（0.723±0.042和0.262±0.037,P＜0.01）.DLC-1 mRNA在正常肝组织、癌旁组织和肝癌组织中的相对表达量分别为0.392±0.094、0.187 ±0.043和0.081 ±0.012,肝癌组织低于正常肝组织（P＜0.05）；DLC-1蛋白在正常肝组织、癌旁组织和肝癌组织中的相对表达量分别为0.423±0.026、0.214±0.029和0.112±0.021,差异有统计学意义（P＜0.01）.miR-181b抑制剂转染SMMC7721细胞后,DLC-1蛋白在空白组、转染组和阴性对照组的相对表达量分别为0.132±0.027、0.379±0.019和0.125±0.015,转染组较其他两组明显升高（P＜0.05）.结论 miR-181b在肝癌组织中表达上调,其抑制DLC-1的表达可能是肝癌发生的重要机制.Objective To investigate the regulatory effect ofmicroRNA-181b （miR-181b） on deleted in liver cancer-1 （DLC-1） and its role in hepatocarcinogenesis.Methods miR-181b and DLC-1 mRNA expression levels in normal tissue,paraneoplastic tissue,liver cancer tissue and liver cancer cells were detected by using real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）.DLC-1 protein expression was detected by using Western blotting.miR-181b inhibitor was transfected into SMMC7721 cells and the expression changes of DLC-1 mRNA and protein were examined by using FQ-PCR and Western blotting respectively.Results miR-181b was up-regulated in hepatocellular carcinoma and adjacent tissues as compared to normal liver tissues and the expression of miR-181 b was also increased in SMMC7721 cells as compared to fetal liver cells LO2,whereas the expression levels of DLC-1 mRNA and protein were decreased in paraneoplastic and HCC tissues compared with normal liver tissues.The expression of miR-181b and DLC-1 was negatively correlated.No obvious difference was found in the DLC-1 mRNA expression after miR-181b inhibitor transfection.Conclusion The DLC-1 abnormal low-expression may play a dynamic role in hepatocarcinogenesis due to the miR-181 b up-regulation.

关 键 词：癌 肝细胞 微小RNA-181b 肝癌缺失基因-1 

分 类 号：R735.7[医药卫生—肿瘤]