| Project/Area Number |
09670566
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | THIRD DEPARTMENT OF INTERNAL MEDICINE, OSAKA CITY UNIVERSITY MEDICAL SCHOOL |
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Principal Investigator |
SHIOMI Susumu OSAKA CITY UNIVERSITY MEDICAL SCHOOL, THIRD DEPARTMENT OF INTERNAL MEDICINE, LECTURER, 医学部, 講師 (30170848)
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| Co-Investigator(Kenkyū-buntansha) |
TAMORI Akihiro OSAKA CITY UNIVERSITY MEDICAL SCHOOL, THIRD DEPARTMENT OF INTERNAL MEDICINE, LECTURER, 医学部, 助手 (30291595)
NISHIGUCHI Shuhei OSAKA CITY UNIVERSITY MEDICAL SCHOOL, THIRD DEPARTMENT OF INTERNAL MEDICINE, ASSOCIATE PROFESSOR, 医学部, 助教授 (10192246)
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| Project Period (FY) |
1997 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | ORNITHINE DECARBOXYLASE / DFMO / HepG2 cell / DIFFERNTIAL DISPLAY / POLYAMINE / PUTRESCINE / cytochrome oxidase / Hep G2細胞 / Cytochrome oxidase / Differential Display法 / ODC遺伝子 / Nuclease Protection Assay / ポリアミン代謝 / Differential dysplay法 / PEST領域 / SSCP法 / ODC過剰発現細胞 / Chang liver cell |
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| Research Abstract |
In order to identify gene differentially expressed by putrescine, a polyamine, which play important roles in the regulation of cell proliferation and the development of cancer, we performed mRNA differential display analysis using total RNA extracted from HepG2 cells (human hepatoblastoma cell line) treated with a specific inhibitor of polyamine biosynthesis, α-difluoromethyl ornithine (DFMO). A total of 25 genes were up-regulated and 32 genes down-regulated by putrescine. Of the genes differentially expressed by putrescine, we chose three that were related to the respiratory chain and oxidative phosphorylase and analyzed them by Northern blot analysis. Cytichrome oxidase submit 1, low molecular mass ubiquinone- binding protein, and NADH dehydrogenase subunit 2 were found to down- regulated by putrescine. We examined intracellular ATP level in Hep G2 cells, and found that ATP level in DFMO- treated cells was increased by exogenous putrescine.
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