大鼠肝癌细胞与胚胎肝细胞microRNA差异表达谱的研究  

Differential expression profile of microRNAs between hepatocellular carcinoma and embryonic hepatocyte in rats

作　　者：吴印涛[1] 李韧[1] 刘卫辉[1] 张福琴[1] 窦科峰[1] 

机构地区：[1]第四军医大学西京医院肝胆外科,西安710032

出　　处：《中华肝胆外科杂志》2009年第12期917-920,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金（30772102）

摘　　要：目的建立大鼠肝癌细胞与大鼠胚胎肝细胞microRNA（miRNA）表达的差异谱，为研究miRNA在肝细胞癌变发生机制中的作用提供新线索。方法化学致癌剂二乙基亚硝胺（DEN）建立近交系F334大鼠肝癌模型，体外原代培养大鼠肝癌细胞和正常F334大鼠胚胎肝细胞，TRIzol法抽提细胞总RNA，进一步制备Hy3荧光标记的miRNA；利用ExiqonmiRNA基因芯片技术，采用含2056条探针的哺乳动物miRNA表达谱芯片对两组之间差异表达的miRNA进行分析，其实验数据用SAM3．0软件进行差异显著性分析。结果原代培养可获得稳定传代的肝癌细胞及胎肝细胞，两者在形态上无明显差异；miRNA芯片共获得78个差异表达的miRNAs，其中68个表达E调（foldchange〉2），10个表达下调（foldchanged 0．5）．部分miRNAs与肿瘤关系密切，其中hsa-miR-21为肝癌癌变相关miRNA。结论正常肝细胞中存在着与肝细胞癌变及侵袭、转移相关的miRNAs，miRNAs的表达具有明显的时序性，其表达特征的改变可能导致了肝癌的发生。Objective To explore the miRNA （rnicroRNA） differential expression profile between hepatocellular carcinoma and embryonic hepatocyte in the rats to provide new evidence that miR NAs are involved in the molecular pathogensis of HCC. Methods The liver cancer model was established by low dose DEN in the rats. Then the hepatocellular carcinoma and embryonic hepatocytes were cultured by primary culture in vitro. The total RNAs were extracted by Trizol and the miRNAs were stained by Hy3. Hybridization was performed by applying the cell miRNAs to mammalian miRNA chip. Data analysis was conducted by using software of SAM of version 3.0. Results A total of 78 differential expressed miRNAs were found and they included 68 over-expression and 10 low-expres sion miRNAs. Data analysis revealed that some miRNAs were associated with some different tumors, especially the has miR-21. Conclusion There are some miRNAs being related to hepatocellular carcinogenesis and invasion and metastasis in the normal hepatocytes. The regularity of the orderly miRNA expression is obvious and the change of expression characteristics of miRNAs may be involved in the pathogensis of HCC.

关 键 词：癌 肝细胞 MIRNA芯片 胎肝细胞 MircoRNAs 

分 类 号：R735.7[医药卫生—肿瘤]