Wang Di-Yi, Zou Li-Ping, Liu Xiao-Jia, Zhu Hong-Guang, Zhu Rong
Department of Pathology, Affiliated Hospital of Taishan Medical University, Taian, 271000, China.
Department of Pathology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
Pathol Oncol Res. 2016 Apr;22(2):393-9. doi: 10.1007/s12253-015-0014-9. Epub 2015 Nov 18.
The hepatitis B virus X protein (HBx), which is encoded by hepatitis B virus (HBV), plays crucial roles in the tumorigenesis of HBV associated hepatocellular carcinoma (HCC). Recent studies suggest that the HBx is involved in regulation of host immune cytokines and chemokines in HBV-associated HCC patients. However, effects of the HBx on autocrine chemokine expression profiles of hepatoma cells, which were shown in modulation of tumor-immune cell interactions, have not been investigated comprehensively. In the present study, human hepatoma cell lines SMMC-7721 and HepG2 were transfected with HBx-expressing plasmid. Human chemokine antibody array 1 (RayBio®), which simultaneously detects 38 chemokine factors, was used to determine chemokine expression profiles. Real-time polymerase chain reaction (real-time PCR) was used to further confirm the differential expression of chemokines. Chemokine antibody array revealed that all 38 chomekines were found to be expressed by SMMC-7721 and HepG2 cell lines. Interleukin-8 (IL-8) was obviously up-regulated, and epithelial neutrophil-activating protein 78 (ENA78), eosinophil chemotactic protein-1 (Eotaxin-1), monocyte chemotactic protein-1 (MCP-1), MCP-2, MCP-3 and macrophage inflammatory protein-3β (MIP-3β) were significantly declined in both cell lines following transfection of HBx-expressing plasmid. Other chemokines showed little or no significant changes. HBx-induced differential chemokine expression levels were validated by real-time PCR. Hierarchical cluster analysis identified a distinction of chomekine expression profiles between HBX-expressing hepatoma cell lines and controls. Our findings provide new evidence that HBx is able to selectively regulate chomekines in hepatoma cells that may be involved in the regulation of tumor-immune cell interactions.
乙型肝炎病毒X蛋白(HBx)由乙型肝炎病毒(HBV)编码,在HBV相关肝细胞癌(HCC)的肿瘤发生中起关键作用。最近的研究表明,HBx参与了HBV相关HCC患者宿主免疫细胞因子和趋化因子的调节。然而,HBx对肝癌细胞自分泌趋化因子表达谱的影响尚未得到全面研究,而这种影响在肿瘤-免疫细胞相互作用的调节中有所体现。在本研究中,用表达HBx的质粒转染人肝癌细胞系SMMC-7721和HepG2。使用能同时检测38种趋化因子的人趋化因子抗体芯片1(RayBio®)来确定趋化因子表达谱。采用实时聚合酶链反应(实时PCR)进一步确认趋化因子的差异表达。趋化因子抗体芯片显示,SMMC-7721和HepG2细胞系均表达所有38种趋化因子。白细胞介素-8(IL-8)明显上调,而上皮中性粒细胞激活蛋白78(ENA78)、嗜酸性粒细胞趋化蛋白-1(嗜酸性粒细胞趋化因子-1)、单核细胞趋化蛋白-1(MCP-1)、MCP-2、MCP-3和巨噬细胞炎性蛋白-3β(MIP-3β)在转染表达HBx的质粒后在两种细胞系中均显著下降。其他趋化因子变化很小或无显著变化。通过实时PCR验证了HBx诱导的趋化因子差异表达水平。层次聚类分析确定了表达HBX的肝癌细胞系与对照之间趋化因子表达谱的差异。我们的研究结果提供了新的证据,表明HBx能够选择性调节肝癌细胞中的趋化因子,这可能参与肿瘤-免疫细胞相互作用的调节。