Identification of hepatitis B virus X-interactive proteins in human hepatocellular carcinoma cell lines HepG2 by using proteomics technique / 中华传染病杂志

ABSTRACT

Objective To identify hepatitis B virus X-interactive proteins by comparative proteomics method and to understand the molecular mechanism of HBx in the development of hepatocellular carcinoma(HCC).Methods Two-dimensional gel electrophoresis(2-DE)was used to separate the total proteins of HBx-transfected human hepatoma cell lines HepG2-Px and its parental cell lines HepG2-P_0.PDQuest software was applied to analyze 2 DE images.The differentially expressed protein spots between the two cell lines were identified by matrix-assisted laser desorptionionization time of flight mass spectrometry(MALDI-TOF-MS)and electrospray ionization tandem mass spectrometry(ESI-Q-TOF).Then,the differential expression levels of some identified proteins were determined by Western blot.The data were compared using t test.Results The well-resolved,reproducible 2-DE patterns of HepG2-Px and HepG2-P_0 total proteins were established.A total of 32 differential proteins were identified in HepG2-Px cell,including 25 up-regulated proteins,such as heat shock protein(HSP)90AB1,Bcl-2 associated athanogene(BAG)-2,nucleophosmin(B23),chloride intracellular channel(CLIC)-1,matrix metalloproteinase(MMP)-3,melanoma antigen(MAGE)-12,and 7 down-regulated proteins,such as Wnt-5a.The differential expression levels of some proteins between the two cell lines were confirmed by Western blot analysis.Conclusions Most of the identified proteins are involved in many processes,such as transcription,signal transduction,cell proliferation,cell cycle regulator,apoptosis,DNA repair,metabolisms and immunity.These differential proteins may play a role in tumor genesis and HC development.The data are valuable for further study on the role of HBx in human hepatocellular carcinoma.