title: [Establishment of an HHCC cell line stably transfected by tumor antigen MAGE-3 gene and its mRNA expression].
abstract: OBJECTIVE
  To construct the eukaryotic expression vector of tumor antigen MAGE-3 and establish human hepatocellular carcinoma cell line (HHCC) expressing MAGE-3.
  
  
  METHODS
  The MAGE-3 gene was amplified by PCR and cloned into the eukaryotic expression vector pIRES2-EGFP to construct the pIRES2-EGFP-MAGE-3 plasmid. The recombinant plasmid pIRES2-EGFP-MAGE-3 was transfected into HHCC cells by lipofectamine, and then the positive clones were screened by G418. The expression of enhanced green fluorescent protein (EGFP) and MAGE-3 mRNA in positive clones were detected by fluorescence microscope and RT-PCR, respectively.
  
  
  RESULTS
  The eukaryotic expression vector pIRES2-EGFP-MAGE-3 was successfully constructed. The expression of EGFP was found by fluorescence microscope detection and MAGE-3 mRNA transcription was detected by RT-PCR in the positive clones.
  
  
  CONCLUSIONS
  The stable MAGE-3-transfected HHCC cell line is successfully established, which will provide experimental basis for further study on immunotherapy for hepatocellular carcinoma using MAGE-3 as target antigen.
authors.affiliation: Department of Pathology, Fourth Military Medical University, Xi'an 710032, China. zhangxm@fmmu.edu.cn
authors.forename: Xiu-min
authors.initials: XM
authors.lastname: Zhang
authors.forename: Yan-fang
authors.initials: YF
authors.lastname: Sui
authors.forename: Shao-yan
authors.initials: SY
authors.lastname: Si
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.forename: Yang
authors.initials: Y
authors.lastname: Huang
authors.forename: Pei-zhen
authors.initials: PZ
authors.lastname: Hu
authors.forename: Wei
authors.initials: W
authors.lastname: Ge
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics*; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000276:immunology / Q000473:pathology* / Q000628:therapy; D045744:Cell Line, Tumor; D004262:DNA Restriction Enzymes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009363:Neoplasm Proteins / Q000235:genetics*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D000951:Antigens, Neoplasm; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004262:DNA Restriction Enzymes