| Project/Area Number |
09470351
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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 |
Urology
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| Research Institution | Nagoya City University |
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Principal Investigator |
KOHRI Kenjiro (1998) Nagoya City University Medical School, chairman, professor, 医学部, 教授 (30122047)
藤田 圭治 (1997) 名古屋市立大学, 医学部, 助手 (50264734)
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| Co-Investigator(Kenkyū-buntansha) |
SASAKI Shoichi Nagoya City University, assistant professor, 医学部, 講師 (50225869)
HAYASHI Yutaro Nagoya City University, assistant professor, 医学部, 講師 (40238134)
坂倉 毅 名古屋市立大学, 医学部, 助手 (00275132)
郡 健二郎 名古屋市立大学, 医学部, 教授 (30122047)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 1998: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1997: ¥11,400,000 (Direct Cost: ¥11,400,000)
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| Keywords | urolithiasis / osteopontin / gene transfection / gene therapy / antisense / NFкBデコイ / ビスホスホネート |
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| Research Abstract |
Urinazy stones contain 1-5% protein, and many reports have suggested the importance of proteins in stone formation. We cloned and sequenced the cDNA encoding osteopontin (OPN), important soluble stone protein components of calcium oxalate stone proteins extracted with O.1M EDTA.Our previous study revealed that osteopontin (OPN), a secreted adhesive phosphoglycoprotein, were strongly expressed by renal distal tubular cells in the rat stone- forming kidney. OPN is clearly produced in the kidney, although its function and significance there remain unclear. Moreover, the association of crystals with renal tubular cells is considered a potential factor in the process of renal stone formation. The mechanisms, however, are not yet well understood. As we spaculated that OPN played a role in crystal-cell interaction, we tried to block the production of OPN by using normal rat kidney cell line (NRK-52E) transfected antisense OPN RNA.The Tetracycline-Regulated Expression System was used. Plasmid PTet-OPNas was constructed by cloning mouse OPN/2ar cDNA sequence in an inverted (antisense) orientation. Two clones of antisense transfectants were identified which showed marked reduction in OPN synthesis upon the absence of tetracycline. Intact NRK-52E cells, NRK-52E cells transfected with empty vector, and tetracycline-treated antisense clones, which did not show inhibition of OPN expression, all under identical conditions associated with calcium oxalate (CaOx) crystals. In contrast, the expression of antisense OPN12ar RNA prevents the association of CaOx crystals with >NRK-52E cells. We suggest that OPN plays a crucial role in this process.
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