Basu Arnab, Meyer Keith, Lai Keith K, Saito Kousuke, Di Bisceglie Adrian M, Grosso Leonard E, Ray Ratna B, Ray Ranjit
Department of Internal Medicine, Liver Center and Cancer Center, Saint Louis University, St. Louis, MO 63110, USA.
Virology. 2006 Jun 5;349(2):347-58. doi: 10.1016/j.virol.2006.02.023. Epub 2006 Mar 20.
Hepatitis C virus (HCV) infection is a major contributor to the development of end-stage liver disease, including cirrhosis and hepatocellular carcinoma (HCC). We have previously shown that HCV core protein promotes immortalization of primary human hepatocytes. To identify molecular changes involved in core protein-mediated immortalization, we have investigated differential gene expression by microarray analyses in primary human hepatocytes and HCV core gene introduced hepatocytes after senescence (early passage), immortalization (middle passage), and anchor-independent growth (late passage). Out of 33,000 human genes screened, 1918 transcripts were differentially expressed (>2-fold) in immortalized human hepatocytes (IHH) as compared to negative controls. Our analyses provided a molecular portrait of changes in gene expression associated with three distinct stages of hepatocytes after introduction of HCV core gene. Many of the overall changes were involved with important cellular pathways, including cell growth regulation, immune regulation, oxidative stress, and apoptosis. We focused on the Stat3 signaling pathway by further verifying selected genes at the protein level relevant to hepatocyte growth regulation. Our data suggested that the introduction of HCV core protein results in an increase in expression of IL-6, gp130, leptin receptor, and Stat3. Upregulation of these genes in turn may regulate c-myc and cyclin D1, downstream of the Stat3 signaling pathway. Identification of these modulated genes with potential roles may help in the selection of targets for therapies against HCV-mediated liver disease progression.
丙型肝炎病毒（HCV）感染是导致终末期肝病（包括肝硬化和肝细胞癌（HCC））的主要因素。我们之前已经表明，HCV核心蛋白可促进原代人肝细胞永生化。为了确定参与核心蛋白介导的永生化过程的分子变化，我们通过微阵列分析研究了原代人肝细胞以及导入HCV核心基因后的肝细胞在衰老（早期传代）、永生化（中期传代）和不依赖贴壁生长（晚期传代）阶段的基因表达差异。在筛选的33000个人类基因中，与阴性对照相比，1918个转录本在永生化人肝细胞（IHH）中差异表达（>2倍）。我们的分析提供了导入HCV核心基因后肝细胞三个不同阶段相关基因表达变化的分子概况。许多整体变化涉及重要的细胞途径，包括细胞生长调节、免疫调节、氧化应激和细胞凋亡。我们通过在蛋白质水平进一步验证与肝细胞生长调节相关的选定基因，重点研究了Stat3信号通路。我们的数据表明，HCV核心蛋白的导入导致IL-6、gp130、瘦素受体和Stat3的表达增加。这些基因的上调反过来可能调节Stat3信号通路下游的c-myc和细胞周期蛋白D1。鉴定这些具有潜在作用的受调控基因可能有助于选择针对HCV介导的肝病进展的治疗靶点。
