Construction of MAGE - 3 Expressing Plasmid for Development of DNA Vaccine Encoding MAGE - 3 Cancer Antigen.
- Author:
Jong Wook PARK
;
Mi Hyun LEE
;
Soo Jung YOON
;
Won Ki BAEK
;
Seong Il SUH
;
Min Ho SUH
;
Kang Dae LEE
;
Tae Hyun YU
- Publication Type:Original Article
- Keywords:
Melanoma antigen gene;
Plasmid;
Vaccine
- MeSH:
Cell Line;
Clone Cells;
DNA*;
DNA, Complementary;
Gene Expression;
Humans;
Immunoblotting;
Immunohistochemistry;
Melanoma;
Plasmids*;
RNA, Messenger;
T-Lymphocytes, Cytotoxic
- From:Journal of the Korean Cancer Association
2000;32(1):191-199
- CountryRepublic of Korea
- Language:Korean
-
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.
