表达HBV的肝癌细胞对巨噬细胞分化影响的初步研究

发布时间：2018-02-19 23:50

本文关键词： 乙型肝炎病毒肝细胞癌巨噬细胞分化基因芯片　出处：《中国病原生物学杂志》2017年10期 　论文类型：期刊论文

【摘要】：目的了解表达乙型肝炎病毒(HBV)的肝癌细胞在体外环境下对巨噬细胞分化的影响。方法收集表达HBV的肝癌细胞HepG2.215和对照肝癌细胞HepG2培养上清,联合佛波酯(PMA,50ng/ml)刺激单核细胞THP-1分化为巨噬细胞,采用Real-time PCR(RT-PCR)检测M1型巨噬细胞分化标志HLA-DR、CD86和M2型巨噬细胞分化标志CD206、CD163的表达;采用基因芯片技术检测分化的巨噬细胞基因表达变化结果表达HBV的HepG2.215细胞培养上清联合PMA成功刺激THP-1分化成巨噬细胞。RT-PCR检测HepG2.215培养上清诱导的巨噬细胞CD163和CD206基因表达上调,HLA-DR基因表达小调;基因芯片的检测显示,与HepG2+PMA比较,HepG2.215培养上清+PMA诱导形成的巨噬细胞有23个基因表达存在差异,其中6个基因表达上调,其余基因表达下调,这些表达差异的基因与基因的转录激活、信号转导、细胞代谢和免疫抑制等生物学功能相关。结论表达HBV的肝癌细胞能诱导巨噬细胞向M2型分化,且多种基因的表达异常与HBV感染肝癌细胞诱导分化的巨噬细胞有关。
[Abstract]:Objective to investigate the effect of hepatoma cells expressing hepatitis B virus (HBV) on macrophage differentiation in vitro. Methods the supernatants of HepG2.215 cells expressing HBV and HepG2 cells of control liver cancer cells were collected. Combined with PMA 50 ng / ml, monocytes were induced to differentiate into macrophages by THP-1. The expression of HLA-DR CD86 and CD206 CD163 was detected by Real-time PCR RT-PCR. Detection of gene expression in differentiated macrophages by gene microarray the supernatants of HepG2.215 cells expressing HBV and PMA successfully stimulated the differentiation of THP-1 into macrophages. RT-PCR was used to detect the CD163 and CD206 bases induced by the supernatants of HepG2.215 culture. The expression of HLA-DR gene was up-regulated in minor; Gene microarray analysis showed that 23 of the macrophages induced by PMA in the supernatant of HepG2.215 were different in gene expression, 6 of which were up-regulated, and the other were down-regulated, compared with HepG2 PMA. These differentially expressed genes are related to the biological functions of transcriptional activation, signal transduction, cell metabolism and immunosuppression. Conclusion hepatoma cells expressing HBV can induce macrophages to differentiate into M2 type. The abnormal expression of many genes was related to macrophages induced by HBV infection in hepatoma cells.
【作者单位】：徐州医科大学病原生物学与免疫学教研室江苏省免疫与代谢重点实验室;江苏省扬州中学高二(8)班;
【基金】：江苏省教育厅高校自然科学研究基金项目(No.16KJB310017) 中国博士后基金项目(No.2016M591925) 江苏省博士后科研资助计划项目(No.1601190C) 徐州医学院课题项目(No.2012KJ13) 江苏高校优势学科建设工程项目(No.PAPD)
【分类号】：R512.62;R735.7

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