MicroRNA在HepG_2肝癌细胞表达差异谱的研究

MicroRNA differential expression profile in the HCC cell lines

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 explore the miRNA （microRNA） differential expression profile between HepG2 and LO2 cell lines, and to provide the evidence that miRNAs are involved in the molecular pathogensis of HCC. Methods HepG2 and LO2 cell total RNAs were extracted by Trizol, then miRNAs were isolated by Ambion＇s miRNA Isolation Kit. Hybridizations were made by applying the cell miRNAs to mammalian miRNA chip. Data analysis was proceeded by software of LuxScan3. 0 and SAM version 2. 1. Results Total 146 differential expressed miRNAs were found, including 80 low-expression and 66 over-expression miRNAs. The highest fold change was up to 36. Data analysis revealed that some miRNAs were associated some different tumors, especially, the hsa miR-224 may play a crucial role in HCC. Conclusion The number of low-expression miRNAs is more than the over-expression in HepG2 compared with LO2 cell, which may indicate that the genes expression are more abundant in HepG2 cells, reflecting the activiated metabolism and rapid proliferation in this cancer. Some miRNAs may be involved in the pathogensis of HCC.