Aim: 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.
Conclusion: 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.