Zhu Bin, Wei Xiao-Xia, Wang Tian-Bao, Zhou Yan-Cai, Liu A-Min, Zhang Guang-Wen
Department of Infectious Diseases, The First Affiliated Hospital, Xin Xiang Medical University, 88 Jian Kang Rd., Wei Hui, 453100, People's Republic of China.
Arch Virol. 2015 Aug;160(8):2043-50. doi: 10.1007/s00705-015-2474-3. Epub 2015 Jun 13.
Hepatitis C virus (HCV) is involved in the initiation and progression of liver fibrosis by regulating genes encoding host proteins. However, the underlying mechanism of HCV-induced liver fibrosis is still to be determined. Reverse transcription polymerase chain reaction (RT-PCR) and western blot were performed to investigate the effect of HCV infection on the expression of the cellular microRNA miR-16 and its target genes encoding hepatocyte growth factor (HGF) and Smad7 in patients infected with HCV and in a liver cell line, QSG-7701, transfected with Ad-HCV, a recombinant adenovirus construct for expression of the HCV core protein. Regulation of HGF and Smad7 expression by miR-16 was assessed using luciferase reporter construct assays and miR-16 mimic transfection. Interferon-α (IFN-α) was used to verify the alteration of gene expression induced by HCV in QSG-7701 cells. Here, we found that miR-16 levels were increased in patients with HCV infection and were correlated with HGF and Smad7 expression levels in patients with HCV infection. Furthermore, HGF and Smad7 were predicted by bioinformatics analysis to be targets of miR-16. Upregulation of miR-16 and decreased HGF and Smad7 expression were still shown in QSG-7701 cells infected with Ad-HCV. Additionally, interferon-α (IFN-α) could reverse the changes in gene expression induced by HCV infection. These results suggest that the upregulation of miR-16 expression induced by HCV infection is a novel mechanism that contributes to downregulation of HGF and Smad7 in the development of liver fibrosis.
丙型肝炎病毒（HCV）通过调控编码宿主蛋白的基因参与肝纤维化的起始和进展。然而，HCV诱导肝纤维化的潜在机制仍有待确定。采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法，研究HCV感染对HCV感染患者以及用表达HCV核心蛋白的重组腺病毒构建体Ad-HCV转染的肝细胞系QSG-7701中细胞微小RNA miR-16及其编码肝细胞生长因子（HGF）和Smad7的靶基因表达的影响。使用荧光素酶报告基因构建体检测和miR-16模拟物转染评估miR-16对HGF和Smad7表达的调控作用。用干扰素-α（IFN-α）验证HCV在QSG-7701细胞中诱导的基因表达变化。在此，我们发现HCV感染患者的miR-16水平升高，且与HCV感染患者的HGF和Smad7表达水平相关。此外，通过生物信息学分析预测HGF和Smad7是miR-16的靶标。在感染Ad-HCV的QSG-7701细胞中仍显示miR-16上调以及HGF和Smad7表达降低。此外，干扰素-α（IFN-α）可逆转HCV感染诱导的基因表达变化。这些结果表明，HCV感染诱导的miR-16表达上调是肝纤维化发生过程中导致HGF和Smad7下调的一种新机制。
