Protein alteration of HepG2.2.15 cells induced by iron overload
Protein alteration of HepG2.2.15 cells induced by iron overload
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铁过载诱导的 HepG2.2.15 细胞蛋白质改变
DOI：
10.1002/pmic.201100335
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发表时间：
2012-05-01
期刊：
影响因子：
3.4
通讯作者：
Lu, Haojie
中科院分区：
文献类型：
--
作者：
Fang, Caiyun;Zhao, Chao;Lu, Haojie
B型肝炎可发展为肝细胞癌。体内铁离子可能干扰B型肝炎病毒（HBV）的清除，并参与肿瘤的发生。为探讨铁在HBV相关致病机制中的作用，本研究观察了不同浓度和价态的铁对HepG2.2.15细胞生长和病毒蛋白分泌的影响。HepG2.2.15细胞对铁刺激具有较强的耐受性，铁刺激后细胞培养液中B型肝炎表面抗原的浓度降低。低浓度的铁促进B型肝炎e抗原（HBeAg）的分泌。Fe ~（2+）比Fe ~（3+）对HBeAg分泌的影响更大。同时，采用iTRAQ和LC-MS/MS研究HepG2.2.15细胞中的差异蛋白质谱。差异表达的蛋白质主要涉及应激反应、信号转导、凋亡等。通过Western-blotting验证了4个蛋白质（14-3-3 beta/alpha、VCP、迁移抑制因子和Nup 153），发现与iTRAQ数据一致。有趣的是，核因子κ B（NF κ B B）的核输入及其活性被发现受到铁刺激的HepG 2.2.15细胞中Nup 153减少的影响。结果提示HBV与铁刺激协同作用损伤宿主肝细胞的可能分子机制。
Hepatitis B can progress into hepatocellular carcinoma. Body irons may interfere with the clearance of hepatitis B virus (HBV) and contribute to genesis of tumor. To investigate the role of iron played in HBV-related pathogenesis, here we studied the effect of iron with different concentrations and valence states on growth of HepG2.2.15 cells and secretion of virus proteins. A strong tolerance of HepG2.2.15 cells to iron challenge was found. The concentration of hepatitis B surface antigen in cell culture medium was decreased after iron stimulation. Lower concentrations of iron facilitated hepatitis B e-antigen (HBeAg) secretion. Fe2+ appeared more effective on HBeAg secretion than Fe3+ did. In parallel, the differential protein profiles in HepG2.2.15 cells were studied by iTRAQ and LC-MS/MS. The differentially expressed proteins were mainly involved in stress response, signal transduction, apoptosis, etc. Four proteins (14-3-3 beta/alpha, VCP, migration inhibitory factor, and Nup153) were verified by Western-blotting and found to be consistent with the iTRAQ data. Interestingly, nuclear import of Nuclear factor kappa B (NF kappa B) and its activity were found to be affected by the decreased Nup153 in iron stimulated HepG2.2.15 cells. The results may indicate possible molecular mechanism how the synergism of HBV and iron stimulation damages host liver cells.