人肝癌细胞株与正常肝细胞株MicroRNA差异表达谱研究  

Differential expression profile of microRNAs between human hepatocellular carcinoma cell line and human normal hepatocyte cell line

作　　者：王家祥[1] 韦源[1] 胡倩[1] 刘秋亮[1] 范应中[1] 杨合英[1] 李苏宁[1] 张大[1] 张谦[1] 

机构地区：[1]郑州大学第一附属医院小儿外科郑州大学肿瘤分子外科研究所,450052

出　　处：《中华肝胆外科杂志》2011年第6期492-496,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：本课题受2008年度河南省高校科技创新团队支持计划项目基金资助（2008IRTSTHN005）

摘　　要：目的建立SMMC-7721人肝癌细胞株与CCC-HEL-1人正常肝细胞株MicroRNA（miRNA）差异的表达谱，确定差异的miRNA，为进一步研究miRNA在肝细胞癌变机制中的作用和肝癌的治疗提供新的线索。方法体外培养SMMC-7721人肝癌细胞株和CCC-HEL-1人正常肝细胞株。用TRIzol法提取细胞的总RNA。用紫外吸收测定法和变性琼脂糖凝胶电泳对RNA进行质量检测。用miRNA芯片技术检测细胞株中miRNA表达。用实时PCR对感兴趣的miRNA进行验证。结果芯片筛选结果显示肝癌细胞株中上调2倍以上的miRNA有154个，4倍以上的miRNA有64个，10倍以上的miRNA有26个；下调2倍以上的84个，下调的4倍以上的22个，下调5倍以上的有10个，下调10倍以上的一个。实时PCR验证结果提示hsa—miR205和hsa-let-7f在肝癌中上调2．7和2．3倍。结论SMMC-7721和CCC-HEL-1细胞株之间存在差异性表达的miRNA分子。在建立的两株细胞miRNA差异表达谱中，发现hsa-miR-205和has-let-7f在肝癌中明显上调。Objective To establish the expression profiles of microRNA （miRNA） in SMMC7721 and CCC-HEL-1 cell lines in order to provide new clues to study the mechanism of miRNA function in hepatocellular carcinoma （HCC） and its treatment. Methods SMMC-7721 and CCC-HEL-1 cell lines were cultured in vitro. Total RNAs were extracted by TRIzol, followed by RNA quantifica- tion and quality control. The RNAs were used to detect the expression of miRNA in these two cell lines by miRNA array. The miRNA of interest was then verified by real-time PCR. Results In total, 238 differentially expressed miRNAs were found,including 154 overexpressed and 84 underexpressed miRNAs. 64 miRNAs were upregulated more than 4 times and26 miRNAs were upregulated more than 10 times. 22 miRNAs were downregulated more than 4 times 10 miRNAs weredownregulated more than 5times, and 1 miRNA was downregulated more than 10 times. Real-time PCR validation suggested that has-miR-205 and has-let-7f in liver cancer increased by 2.7 and 2.3 times, respectively. Conclusion There are differences in the expression of miRNA between the SMMC-7721 and CCCHELl cell lines and the profiles of miRNA expression were established for both cell lines. It was found that has-miR-205 and has-let-7f were upregulated in liver cancer.

关 键 词：肝细胞癌 肝细胞 细胞株 小核糖核酸 

分 类 号：R735.7[医药卫生—肿瘤]