1Institute for Medical Science, Keimyung University School of Medicine. 2Institute for Medical Science, College of Medicine, Kosin University, Taegu, Korea.
ABSTRACT
PURPOSE: The spectrum of melanoma antigen gene (MAGE)-expressing tumor is very wide and the gene of MAGE express antigens that are targets for specific recognition by cytotoxic T lymphocytes derived from tumor-bearing patients. All of these characteristics represent MAGE as tumor vaccine can be useful for cancer prevention or treatment. Here, we detected MAGE-3 gene expression in cancer cell lines and evaluated recombinant MAGE-3 protein producibility of MAGE plasmid to develope MAGE DNA vaccine.
MATERIALS AND METHODS: MAGE-3 gene expression of cancer cell lines was evaluated by reverse transcription-polymerase chanin reaction (RT-PCR).
Two kinds of MAGE-3 expressing plasmids were constructed and their MAGE-3 protein producibility was evaluated by immunohistochemistry and immunoblotting using monoclonal anti-MAGE-3 antibody.
RESULTS: Among 13 cell lines, SNU484, AMC-HN-3, AMC-HN-4, AMC-HN-7, HeLa, NCI H1703 and HT29 expressed MAGE-3 mRNA. In order to make MAGE plasmid, cDNA that showed 100% DNA homology with MAGE-3 gene was cloned into pcDNA 3 plasmid and pSecTag plasmid. Intracytoplasmic and secretory recombinant MAGE-3 was produced by MAGE-3 containing pcDNA 3 plasmid and pSecTag plasmid, respectively.
CONCLUSION: In this study, we showed high expression frequency of MAGE-3 in cancer cell line, and established two kinds of plasmid that produce recombinant MAGE-3 in cell lines. We expect these plasmids will be used in cancer treatment or MAGE-3 function study in future.
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