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J Korean Cancer Assoc > Volume 32(1); 2000 > Article
Journal of the Korean Cancer Association 2000;32(1): 191-199.
Construction of MAGE - 3 Expressing Plasmid for Development of DNA Vaccine Encoding MAGE - 3 Cancer Antigen
Jong Wook Park, Mi Hyun Lee, Soo Jung Yoon, Won Ki Baek, Seong Il Suh, Min Ho Suh, Kang Dae Lee, Tae Hyun Yu
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.
Key words: Melanoma antigen gene;Plasmid;Vaccine
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