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目的研究HBX蛋白对肝细胞内mi RNA表达水平的影响。方法构建表达HBX的慢病毒质粒,慢病毒包装后转染L02和Hep G2细胞,通过mi RNA芯片分析筛选出表达变化差异明显的mi RNA,并进一步对mi RNA的靶基因进行预测。结果成功构建了稳定表达HBX的L02-HBX和Hep G2-HBX细胞系,芯片分析发现了19个表达异常的mi RNA,预测到304个最有可能的潜在靶基因,在肝癌细胞系和肝癌组织中,通过RT-PCR验证了mi RNA-21、mi RNA-211和mi RNA-125b及其靶基因BCL2L10和ARHGAP10的异常表达。结论 HBX通过诱导肝细胞内mi RNA表达谱发生改变,导致相关抑癌基因失活或癌基因活化,进而影响HCC的发生发展。
Objective To investigate the effect of HBX on the expression of miRNA in hepatocytes. Methods Lentiviral plasmids expressing HBX were constructed. The lentivirus was packaged and transfected into L02 and Hep G2 cells. The mi RNAs with significant differences in expression were screened by mi RNA microarray. The miRNA target genes were further predicted. RESULTS: L02-HBX and Hep G2-HBX cell lines stably expressing HBX were successfully constructed and 19 miRNAs with abnormal expression were found by microarray analysis. The 304 most probable potential target genes were predicted, and in liver cancer cell lines and liver cancer tissues , Mi RNA-21, mi RNA-211 and mi RNA-125b and their target genes BCL2L10 and ARHGAP10 were verified by RT-PCR. Conclusion HBX induces the change of miRNA expression profile in hepatocytes, resulting in the inactivation of oncogene or oncogene activation, thereby affecting the occurrence and development of HCC.