title: Protein alteration of HepG2.2.15 cells induced by iron overload.
abstract: 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. Fe(2+)  appeared more effective on HBeAg secretion than Fe(3+)  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 β/α, 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 (NFκB) 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.
authors.affiliation: Department of Chemistry, Fudan University, Shanghai, P. R. China.
authors.forename: Caiyun
authors.initials: C
authors.lastname: Fang
authors.forename: Chao
authors.initials: C
authors.lastname: Zhao
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Liu
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
authors.forename: Haojie
authors.initials: H
authors.lastname: Lu
mesh_terms: D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002853:Chromatography, Liquid; D005296:Ferrous Compounds / Q000494:pharmacology; D056945:Hep G2 Cells; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000032:analysis / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D019190:Iron Overload / Q000378:metabolism* / Q000821:virology; D007553:Isotope Labeling; D008113:Liver Neoplasms / Q000378:metabolism / Q000821:virology; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
chemical_list: D005296:Ferrous Compounds; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D016328:NF-kappa B; C079377:NUP153 protein, human; D028861:Nuclear Pore Complex Proteins; D020543:Proteome; C020748:ferrous sulfate