| Project/Area Number |
15590691
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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 | Keio University (2005)
Tokai University (2003-2004) |
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Principal Investigator |
HOZAWA Shigenari Keio University, School of Medicine, Assistant professor, 医学部, 専任講師 (40181482)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Hirotoshi The University of Tokyo, The Institute of Medical Science, Associate professor, 医科学研究所, 助教授 (00171794)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2005: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | hepatic stellate cell / pancreatic stellate cell / rho GTPase family / MMP-1 / NF-kappa B / p50 homodimer / gene transcription / 低分子量G蛋白 / Rhoファミリー / Phoファミリー |
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| Research Abstract |
The aim of this study is to investigate the gene regulation of MMP-1 by rho GTPase family in human hepatic and pancreatic stellate cells, and to identify the transcriptional factor of the gene regulation.
1. The expression of MMP-1 protein, mRNA and its gene transcription were upregulated by IL-1 beta in both hepatic stellate cells and pancreatic stellate cells.
2. Morphological changes were observed in stellate cells by treatment of a specific inhibitor of rho A protein ; C3-transferase. Since the changes were transient and reversible, a rho A protein in the stellate cells was suggested to be functional. C3-transferase did not inhibit the induction of MMP-1 expression by IL-1 beta. The cotransfection studies using dominant-negative mutant plasmids for rho family members and MMP-1 gene promoter plasmids suggested that rac 1 GTPase may regulate the induction of MMP-1 gene expression by IL-1 beta in gene transcriptional level.
3. The deletional analysis and site-directed mutagenesis revealed that both IL-1 beta and rac 1 may upregulate MMP-1 gene promoter activity via a putative NF-kappaB site at -2541 bp of 5'-flanks. Furthermore, gelshift and supershift assay revealed that both TNF-alpha and rac 1 GTPase may regulate MMP-1 gene transcription via a p50 homodimer of NF-kappa B in hepatic stellate cells. In human pancreatic stellate cells, IL-1 beta was suggested to regulate MMP-1 gene transcription via rac 1 GTPase, but not via NF-kappa B.
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