Guan Shi-He, Yang Kai, Lu Meng-Ji, Lu Yin-Ping, Yang Dong-Liang
Department of Laboratory, Second Affiliated Hospital of Anhui Medical University, Hefei 230601, China.
Zhonghua Gan Zang Bing Za Zhi. 2011 Jun;19(6):440-4.
To investigate the possible influence of HBV and its antigens on the expressions of JAK-STAT signal transduction pathway molecules and the antiviral proteins of IFN alpha.
The HepG2 cells were transfected with pSM2, pHBS2-S and pHBc-EGFP plasmids which express HBV whole particles or S-antigen, Pre-S antigen and core antigens. The infectious supernatant from HepG2.2.15 cells and the pured HBV proteins which contained the S, Pre-S antigens were used to treat the HepG2 cells. Northern blot and RT-PCR were applied to analyse the expressions of the antiviral proteins MxA, 2' -5' OAS, 9-27 and the JAK-STAT signal transduction pathway molecules STAT1 in HepG2 cells responded to the IFN alpha treatment.
The HepG2 cells transfected with pSM2, pHBS2-S and pHBc-EGFP plasmids could express whole HBV particles and HBsAg, Pre-S antigen and HBcAg. The quantitation of expressed HBV particles and antigens increased significantly during the course of transfection. Northern blot hybridization analysis indicated that the HepG2 cells expressed IFN alpha antiviral proteins MxA, 2' -5' OAS and 9-27. When transfected with pHBV-dimer, pHBS2-S, pHBc-EGFP plasmids, the IFN/A antiviral proteins MxA, 2' -5' OAS and 9-27 in transfected cells were reduced greatly as compared to the un-transfected HepG2 cells, and the expressed antiviral proteins decreased sharply with the development of transfection time. Furthermore, the expression of IFN alpha JAK-STAT signal transduction pathway molecule STAT1 was also inhibited with the expression of HBV particles and HBV antigens in transfected HepG2 cells.
The HBV and its antigens influence the expressions of IFN alpha JAK-STAT signal transduction pathway molecules and antiviral proteins in the hepatocellular models in vitro. It is indicated that HBV might possess the activity to antagonise or counteract the IFN alpha antiviral action.
研究乙肝病毒（HBV）及其抗原对JAK-STAT信号转导通路分子以及干扰素α（IFNα）抗病毒蛋白表达的可能影响。
用表达HBV完整颗粒或S抗原、前S抗原及核心抗原的pSM2、pHBS2-S和pHBc-EGFP质粒转染HepG2细胞。用HepG2.2.15细胞的感染性上清液以及含S、前S抗原的纯化HBV蛋白处理HepG2细胞。采用Northern印迹法和逆转录-聚合酶链反应（RT-PCR）分析HepG2细胞在IFNα处理后抗病毒蛋白Mx A、2'-5'寡腺苷酸合成酶（OAS）、9-27以及JAK-STAT信号转导通路分子信号转导子和转录激活子1（STAT1）的表达。
用pSM2、pHBS2-S和pHBc-EGFP质粒转染的HepG2细胞可表达完整HBV颗粒以及乙肝表面抗原（HBsAg）、前S抗原和乙肝核心抗原（HBcAg）。转染过程中表达的HBV颗粒和抗原量显著增加。Northern印迹杂交分析表明，HepG2细胞表达IFNα抗病毒蛋白Mx A、2'-5'OAS和9-27。与未转染的HepG2细胞相比，用pHBV-二聚体、pHBS2-S、pHBc-EGFP质粒转染的细胞中，IFN/α抗病毒蛋白Mx A、2'-—5'OAS和9-27显著减少，且随着转染时间的延长，表达的抗病毒蛋白急剧下降。此外，转染的HepG2细胞中，随着HBV颗粒和HBV抗原的表达，IFNα JAK-STAT信号转导通路分子STAT1的表达也受到抑制。
HBV及其抗原在体外肝细胞模型中影响IFNα JAK-STAT信号转导通路分子和抗病毒蛋白的表达。提示HBV可能具有拮抗或抵消IFNα抗病毒作用的活性。
