[Establishment of an HHCC cell line stably transfected by tumor antigen MAGE-3 gene and its mRNA expression].
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.
Zhang
Xiu-min
XM
Department of Pathology, Fourth Military Medical University, Xi'an 710032, China. zhangxm@fmmu.edu.cn
Sui
Yan-fang
YF
Si
Shao-yan
SY
Li
Xia
X
Huang
Yang
Y
Hu
Pei-zhen
PZ
Ge
Wei
W
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
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