HBV病毒X蛋白诱导肝细胞miRNA表达谱变化的研究

Comprehensive analysis of microRNA expression profile in HBX-induced hepatocellular carcinoma

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。

Hepatocellular carcinoma （HCC） is one of the most prevalent malignant diseases worldwide and Hepatic B virus infection is the major risk factor in east-Asia and Sub-Saharan Africa. Hepatitis B virus-encoded X protein （HBX） has been widely reported to play important and extensive roles in the pathogenesis of HCC. And some microRNAs （miRNAs） are getting more attentions in carcinogenesis of HCC. However, the global changes of miRNA expression in hepatocellular induced by HBX protein have been less studied. Here, we transfected the HBx gene into L02 and HepG2 cells using lentivirus, and investigated the influence of HBX protein on the miRNA expression by miRNA microarray assay. We found 19 significantly different expressed miRNAs and predicted 304 potential target genes regulated by these miRNAs. Using quantitative RT-PCR, we identified that the different expression of miRNA-21, miRNA-211 and miRNA-125b in HepG2-HBX cell and HCC tissues was consistent with the results of miRNA microarray and the expression level of these miRNAs target gene also changed. In conclusion, we have comprehensively analyzed the global influence of HBX protein on hepatocyte miRNA expression and found more miRNA targets induced by HBX protein.