乙肝病毒X蛋白上调骨桥蛋白表达增强肝癌细胞干性的研究

发布时间：2018-03-03 02:16

本文选题：骨桥蛋白　切入点：乙肝病毒x蛋白　出处：《免疫学杂志》2017年07期 　论文类型：期刊论文

【摘要】：目的在乙型肝炎病毒(HBV)相关性肝细胞肝癌(hepatocellular carcinoma,HCC)中探讨乙肝病毒x蛋白(HBx)对骨桥蛋白(osteopontin,OPN)表达及肝癌细胞干性的影响。方法采用免疫印迹(Western blot,WB)及免疫组化等方法在肝癌组织标本上检测OPN表达水平与患者性别、年龄、AFP、HBV病毒复制量、肿瘤大小、TNM分期等临床病理因素的关系;在稳定整合HBV基因组的HepG2.2.15细胞系及慢病毒介导的HBx转染HepG2细胞中,进一步验证HBV及HBx过表达对OPN及干性相关转录因子Nanog、Oct4,干性细胞表面标记分子CD44、CD90、Ep CAM表达水平的影响。结果免疫组化及WB检测结果均表明OPN在HBV阳性的HCC肝癌组织中的表达水平显著高于HBV阴性的肝癌组织(P0.05)。HepG2.2.15细胞中OPN的mRNA及蛋白质含量较HepG2细胞显著增加(P0.05);在HepG2细胞中,过表达HBx显著上调OPN mRNA及蛋白质水平(P0.05),流式细胞术定量分析显示过表达HBx的HepG2细胞中Ep CAM、CD90阳性的干性细胞数量明显升高(P0.05),实时定量PCR显示干性相关转录因子Nanog、Oct4及其表面标志分子CD44、CD90、Ep CAM水平显著升高。结论 HBx能够显著上调OPN的表达,并增强肝癌细胞的干性,提示HBx/OPN/肝癌细胞干性增强通路可能在HBV相关HCC患者预后不良中具有重要作用。
[Abstract]:Objective to investigate the effect of hepatitis B virus x protein (HBX) on the expression of osteopontin OPN (OPN) and the dryness of hepatoma cells in hepatocellular carcinoma (HCC) associated with hepatitis B virus (HBV). The expression level of OPN and the sex of patients were detected in cancer tissues. Clinical and pathological factors, such as HBV replication, tumor size, TNM staging, etc., in the HepG2.2.15 cell lines that stably integrate the HBV genome and the lentivirus-mediated HBx transfection into HepG2 cells, To further verify the effect of HBV and HBx overexpression on the expression of OPN and the dry-related transcription factor Nanog-Oct4, CD4hCD90Ep CAM, a marker molecule on the surface of dry cells. Results the results of immunohistochemistry and WB showed that OPN was present in HBV positive HCC HCC tissues. The mRNA and protein contents of OPN in P0.05N. HepG2.2.15 cells were significantly higher than those in HepG2 cells, while in HepG2 cells, the expression level of OPN was significantly higher than that of HepG2 cells. Overexpression of HBx significantly upregulated OPN mRNA and protein level (P0.05). Quantitative analysis by flow cytometry showed that the number of CD90 positive dry cells in HepG2 cells with HBx overexpression was significantly higher than that of P0.05. Real-time quantitative PCR showed the dry related transcription factor Nanogan Oct4, and. The expression of OPN was significantly up-regulated by HBx. The increased dryness of HBx/OPN/ hepatoma cells may play an important role in the poor prognosis of HBV related HCC patients.
【作者单位】：第三军医大学大坪医院野战外科研究所肝胆外科;第三军医大学生物化学与分子生物学教研室;第三军医大学西南医院病理研究所;重庆医科大学分子医学与肿瘤研究中心;重庆第三军医大学大坪医院野战外科研究所肝胆外科;
【基金】：国家自然科学基金(81372270,81270482)
【分类号】：R735.7

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