title: [The different structure forms of HBx protein influence their intracellular distribution in hepatocellular carcinoma HepG2 cells].
abstract: OBJECTIVE
  Different eukaryotic expression vectors were selected in this research, and then we respectively cloned and constructed recombinant plasmids contained HBX gene or its different fusion forms with GFP. We are aimed to explore the influence of the HBX protein with different structure on its intracellular localization.
  
  
  METHODS
  Flag-HBX gene was amplified from pcDNA3.0-HBX plasmid in our laboratory, cloned into pMD-18T vectors, sequenced.and then subcloned into different vectors. After right sequenced, respective recombinant plasmids of pFlag- HBX-IRES2-EGFP, pEGFP-C3-Flag-HBX and pFlag-HBX-EGFP-N3 were transiently transfected into hepatoma HepG-2 cells. The intra-cellular localizations and distributions of HBX protein were examined by indirect immunofluorescence.
  
  
  RESULTS
  Three different Flag-HBX eukaryotic expression vectors were successfully constructed. After transfection of them into HepG2 cells respectively, indirect immunofluorescence demonstrated that HBX protein fused with GFP in different forms show distint intra-cellular distribution characteristics.
  
  
  CONCLUSIONS
  We have provided significant experimental evidences for research of the role of HBX protein in the development of hepatocellular carcinoma, and especially the references for explanation of the outcomes in vitro transfection experiments.
authors.affiliation: Department of Oncology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Peng
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Jie
authors.initials: J
authors.lastname: Min
authors.forename: Chang-yu
authors.initials: CY
authors.lastname: Pu
authors.forename: Hou-long
authors.initials: HL
authors.lastname: Zhou
authors.forename: An-gang
authors.initials: AG
authors.lastname: Yang
authors.forename: Ying-ming
authors.initials: YM
authors.lastname: Feng
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular / Q000379:methods*; D056945:Hep G2 Cells / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D011994:Recombinant Proteins; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein