大鼠肝癌细胞与胚胎肝细胞的microRNA差异表达谱分析  

Differential expression profile of microRNA between hepatocellular carcinoma cells and embryonic hepatocytes in rats

作　　者：吴印涛[1] 邱宝安[1] 

机构地区：[1]中国人民解放军海军总医院,北京100036

出　　处：《山东医药》2013年第37期8-1,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(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Exiqon miRNA基因芯片技术,采用含2 056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。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个表达上调,10个表达下调,部分miRNAs与肿瘤关系密切,其中miR-122为肝细胞癌变相关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,主要参与转录后基因表达的调控,其表达特征的改变可能导致了肝癌的发生。Objective To establish the microRNA( miRNA) differential expression profile between hepatocellular carcinoma( HCC) cells and embryonic hepatocytes in rats,and to provide the evidence that miRNAs are involved in the molecular pathogenesis of HCC. Methods The inbred line F334 rat hepatoma models were established by low dose diethylnitrosamine( DEN),and then embryonic hepatocytes in healthy F334 rats and hepatocellular carcinoma cells were conducted by primary culture in vitro. The total RNAs were extracted by Trizol,and then miRNAs were stained by Hy3. Hybridizations were made by applying the cell miRNAs to mammalian miRNA chip. Data analysis was proceeded by software of SAM version 3. 0. Results The stable passage of HCC cells and embryonic hepatocytes was obtained in primary culture,and with no significant differences in morphology. Totally 78 differential expressed miRNAs were found,including 68 up-regulated and 10 down-regulated miRNAs. Data analysis revealed that some miRNAs were associated with some different tumors,and miR-122 was the miRNA related to HCC. Conclusions There are some miRNAs related to hepatocellular carcinogenesis,invasion and metastasis in the normal hepatocytes. The expression of miRNAs shows obvious regularity and is mainly involved in the regulation of gene expression after transcription,meanwhile,the changes of expression characteristics of miRNAs may lead to HCC.

关 键 词：肝肿瘤 RNA 小核 胎肝细胞 大鼠 

分 类 号：R735.7[医药卫生—肿瘤]