不同来源人肝细胞系β干扰素诱生及其信号通路的比较

Interferon-β induction and related signaling pathways in human hepatocyte cell lines

β干扰素(IFN-β)是一种在抗病毒固有免疫应答过程中具有重要作用的细胞因子,而肝细胞作为肝炎病毒的宿主细胞被认为具有IFN-β诱生的能力,但目前尚未建立合适的体外人肝细胞模型用以研究乙型肝炎病毒(HBV)与肝细胞干扰素系统的相互作用。为此,本研究选取了3株人肝细胞系PH5CH8、Huh7、HepG2作为研究对象,以IFN-β诱生剂新城疫病毒(NDV)与多聚次黄苷酸-胞苷酸〔poly(I∶C)〕处理细胞,从转录、蛋白水平及功能学角度检测产生IFN-β的能力。结果显示,与Huh7和HepG2细胞相比,PH5CH8细胞经NDV与poly(I∶C)诱导可产生高水平的IFN-β。进一步利用蛋白免疫印迹法比较3株细胞系内IFN-β诱生信号通路相关蛋白的表达水平,结果显示,与PH5CH8细胞相比,Huh7和HepG2细胞内多种信号蛋白的表达水平偏低。本研究结果为选择合适肝细胞系用于HBV与干扰素系统作用的研究提供了实验依据,并为重建IFN诱生系统选择性缺陷的肝细胞系提供了理论基础。

Interferon-β plays an important role in innate immunity against viral infection. Hepatocytes, as hepatitis viruses-harboring cells, are reported to possess the potential to inductively express interferon-β. However, a practical in vitro cell model for investigating the interplay between hepatitis B virus and host cells is rarely reported. Here, we determined the inductive expression of interferon-β by interferon-β agonists [ （ Newcastle disease virus, NDV ） and poly （I： C）3 in immortalized primary hepatic cell line, PH5CH8, and hepatocarcinoma cell lines, Huh-7 and HepG2. The data demonstrated that inductive expression of interferon-β in PH5CH8 cells was significantly higher than that in Huh7 or HepG2 cells. In addition, the expression level of key molecules critical for interferon-β induction was investigated to clarify the underlying mechanism. The results showed that the background expression level was fairly low in Huh7 and HepG2 cell lines, compared to that in PH5CH8 cells. It is suggested that PH5CH8 cells possess intact potential to produce interferon-β and reconstitution of a selective interferon-deficient hepatic cell line might be achieved via introduction of related molecules critical for interferon induction.