微RNA-21在肝细胞癌中的表达及其靶基因研究  

Expression and function research of miRNA-21 in hepatocellular carcinoma

作　　者：刘长征[1] 蔡磊[2] 刘卫[2] 杨克恭[1] 何小东[2] 陈松森[1] 

机构地区：[1]中国医学科学院北京协和医学院基础医学研究所医学分子生物学国家重点实验室,100005 [2]北京协和医院

出　　处：《中华实验外科杂志》2009年第8期964-966,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨微RNA（miRNA）．21在肝细胞癌中的表达特征及其功能，筛选并鉴定miRNA-21可能调控的靶基因。方法用PAGE／Northern blot方法分析miRNA-21在10例肝细胞癌患者组织标本中的表达特征，并在3种肝癌细胞株中加以验证。用生物信息学分析预测miRNA-21可能的靶基因，通过逆转录一聚合酶链反应（RT—PCR）方法克隆靶基因的调控区序列，利用荧光素酶双报告基因系统筛选，并通过Western blot、流式细胞仪在蛋白质水平进行分析验证。用体外细胞侵袭实验分析miRNA-21与肝癌细胞侵袭能力的关系。结果miRNA-21在10例肝细胞癌患者中均表达上调，与癌旁组织比较，表达量增加10倍以上。在3种肝癌细胞株中也得到相似的结果。筛选并确认了RECK为miRNA-21的调控靶基因之一。抑制miRNA-21的表达可明显降低肝癌细胞的侵袭。结论miRNA-21在肝细胞癌中表达失控，通过抑制RECK表达促进肿瘤细胞侵袭。Objective To research the expression signature and function of miRNA-21 in hepatocellular carcinoma （ HCC）, and indentify its functional target mRNA. Methods The expression signature of miRNA-21 in 10 cases of HCC was confirmed in 3 kinds of HCC cell lines by PAGE/Northern blot. The potential target mRNA prediction of miRNA-21 was performed by bioinformatics. The regulation region of miRNA-21 in RECK mRNA was cloned by RT-PCR, and confirmed by sequencing. Dual report gene assay was used to identify the target gene of miRNA-21. The target gene was confirmed by Western blot and FCM analysis. The effect of miRNA-21 on HCC cell line HepG2 was analyzed by in vitro invasion assay. Results miRNA-21 was over-expressed in 10 cases of HCC and 3 kinds of cell lines. RECK was identified as one of miRNA-21 target genes. The invasion ability of HepG2 was suppressed when miRNA-21 was knocked down. Conclusion miRNA-21 was over-expressed in HCC, and promoted cancer cell invasion by inhibiting the expression level of RECK.

关 键 词：癌 肝细胞 MIRNA RECK 侵袭 

分 类 号：R735.7[医药卫生—肿瘤] R394.8[医药卫生—临床医学]