Interferon-α response in chronic hepatitis B-transfected HepG2.2.15 cells is partially restored by lamivudine treatment
Interferon-α response in chronic hepatitis B-transfected HepG2.2.15 cells is partially restored by lamivudine treatment
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DOI：
10.3748/wjg.v13.i2.228
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发表时间：
2007-01-14
影响因子：
4.3
通讯作者：
Schlaak, Joerg F.
中科院分区：
文献类型：
--
作者：
Guan, Shi-He;Lu, Mengji;Schlaak, Joerg F.
目的：研究抗病毒化合物拉米夫定对乙肝病毒感染的HepG2.2.15细胞的干扰素应答及其调节作用。方法：用不同浓度的干扰素-α2a刺激HepG2.2.15和HepG2细胞。然后，提取总RNA，并通过定制的cDNA阵列和干扰素诱导基因(ISGs)的Northern杂交进行分析。此外，提取细胞蛋白用于EMSA和Western印迹。结果：在HepG2.2.15细胞中发现2个基因(mxa、Cig5)完全缺失，4个基因(IFITM1、-2、-3、6-16)部分降低干扰素应答。在2个基因(IFITM1，6-16)中，拉米夫定可恢复对干扰素-α的应答。这种作用不能通过直接调节JAK/STAT信号通路来解释，因为EMSA和Western印迹实验显示，与HepG2细胞相比，经干扰素-α刺激后，HepG2.2.15中的STAT1激活和ISGF3的形成没有受到抑制。结论：这些结果与慢性乙型肝炎可能通过选择性地阻断某些ISG来特异性地调节细胞对干扰素的反应的假设是一致的。拉米夫定的抗病毒治疗可能通过减少乙肝病毒基因表达和复制来部分恢复ISG的表达。(C)2006年《假发出版社》。版权所有。
AIM: To characterize the IFN-response and its modulation by the antiviral compound lamivudine in HBV-transfected HepG2.2.15 cells.METHODS: HepG2.2.15 and HepG2 cells were stimulated with various concentrations of IFN-alpha 2a in the presence or absence of lamivudine. Then, total RNA was extracted and analysed by customised cDNA arrays and northern blot for interferon-inducible genes (ISGs). In addition, cellular proteins were extracted for EMSA and western blot. HBV replication was assessed by southern blot or ELISAs for HBsAg and HBeAg.RESULTS: Two genes (MxA, Cig5) with completely abolished and 4 genes (IFITM1, -2, -3, and 6-16) with partially reduced IFN-responses were identified in HepG2.2.15 cells. In 2 genes (IFITM1, 6-16), the response to IFN-alpha could be restored by treatment with lamivudine. This effect could not be explained by a direct modulation of the Jak/Stat signalling pathway since EMSA and western blot experiments revealed no suppression of Stat1 activation and ISGF3 formation after stimulation with IFN-alpha in HepG2.2.15 compared to HepG2 cells.CONCLUSION: These results are consistent with the assumption that chronic hepatitis B may specifically modulate the cellular response to IFN by a selective blockage of some ISGs. Antiviral treatment with lamivudine may partially restore ISG expression by reducing HBV gene expression and replication. (c) 2006 The WIG Press. All rights reserved.