2004 Fiscal Year Final Research Report Summary
Prechnical research on gene therapy for oral cancer using a RAS dominant negative mutant
| Project/Area Number |
14370651
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Surgical dentistry
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KUZUMAKI Noboru Hokkaido University, Institute for Genetic Medicine, Prof., 遺伝子病制御研究所, 教授 (80091445)
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| Co-Investigator(Kenkyū-buntansha) |
CHIBA Itsuo Health Sciences University of Hokkaido, Graduate School of Dentistry, Prof., 歯学研究科, 教授 (50250460)
TAKIMOTO Masato Institute for Genetic Medicine, Asso, Prof., 遺伝子病制御研究所, 助教授 (30179585)
NAKAGAWA Koji Institute for Genetic Medicine, Lec., 遺伝子病制御研究所, 助手 (80360949)
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| Project Period (FY) |
2002 – 2004
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| Keywords | RAS oncogene / suppressive mutant / tongue cancer / growth suppression / telomerase |
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| Research Abstract |
In oral cancers, the abnormal expression of RAS oncogene is a critical event in tumor growth and metastasis. This study was performed to test the efficacy of a dominant negative RAS mutant, N116Y, in blocking the growth of human tongue cancer cell lines using a replication-deficient recombinant adenoviral vector as basic experiments for gene therapy. Infection with N116Y adenovirus (Ad.CMV-N116Y) but not with LacZ adenovirus (Ad.CMV-LacZ) in which the expression was driven by the cytomegalovirus promoter, significantly reduced the in vitro growth and induced abnormal morphology and apoptotic picnosis of the tongue cancer cell lines (HSC-3, HSC-4, SAS) studied. To examine the suppressive mechanism of N116Y, the activation of extracellular-signal regulated kinase (ERR) were examined by Western blot analysis. Infection with Ad.CMV-N116Y suppressed the activation of ERK after EGF stimulation in serum-starved HSC-3 cells. To evaluate the tumor specific cytotoxicity of N116Y for suppressing growth of tongue cancer cell lines, we also constructed a replication deficient recombinant N116Y adenovirus vector driven by human telomerase reverse transcriptase (hTERT) promoter (Ad.hTERT-N116Y) or a control virus vector Ad.hTERT and showed tongue cancer-specific cytotoxicity by Ad.hTERT-N116Y but not by Ad.hTERT using the MTT assay. These findings indicate that N116Y is a potential candidate gene for human tongue cancer gene therapy.
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