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目的建立HepG2人肝癌细胞株与LO2正常肝细胞株microRNA(miRNA)表达的差异谱,为研究miRNA在肝细胞癌变机制中的作用提供新线索。方法体外培养HepG2人肝癌细胞和LO2人正常肝上皮细胞,Trizol法抽提细胞总RNA,Ambion′s miRNA Isolation Kit进一步分离miRNA;利用基因芯片技术,将细胞miRNA与哺乳动物miRNA芯片杂交,采用Lux-Scan3.0图像软件和SAM version 2.1进行数据分析。结果HepG2细胞与LO2细胞表达的miRNA中有146个存在着明显差异(fold change>4.0),其中80个低表达和66个高表达,最高fold change达36倍,部分miRNAs与肿瘤关系密切,其中has-miR-224为肝癌癌变相关miRNA。结论HepG2较LO2细胞miRNA低表达数多于高表达数,反映其基因表达数量更丰富,细胞代谢和增殖更活跃;部分miRNAs可能参与肝细胞癌变分子机制。
Objective To establish a differential spectrum of microRNA (miRNA) expression in HepG2 human hepatoma cell line and normal LO2 cell line, providing new clues for studying the role of miRNA in hepatocellular carcinogenesis. Methods HepG2 human hepatocellular carcinoma cells and LO2 human normal liver epithelial cells were cultured in vitro. Total RNA was extracted by Trizol method and further separated by Ambion’s miRNA Isolation Kit. The miRNAs were hybridized with mammalian miRNAs by using gene chip technology. -Scan3.0 image software and SAM version 2.1 for data analysis. RESULTS: There were 146 miRNAs in HepG2 cells and LO2 cells with fold change> 4.0. Among them, 80 were low expression and 66 were high expression, the highest fold change was 36-fold. Some miRNAs were closely related to tumor, has-miR-224 is a cancer-associated miRNA for hepatocellular carcinoma. Conclusion Compared with LO2 cells, HepG2 cells express more miRNAs than those with high expression levels, reflecting more abundant gene expression and more active cell metabolism and proliferation. Some miRNAs may be involved in the molecular mechanism of hepatocellular carcinoma.