| Project/Area Number |
15591193
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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 |
Dermatology
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| Research Institution | Keio University |
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Principal Investigator |
MATSUZAKI Yuriko Keio University, Department of Medicine, Instructor, 医学部, 助手 (40255435)
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| Co-Investigator(Kenkyū-buntansha) |
SUMIMOTO Hidetoshi Keio University, Department of Medicine, Instructor, 医学部, 助手 (00306838)
KAWAKAMI Yutaka Keio University, Department of Medicine, Professor, 医学部, 教授 (50161287)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | RNA interference / melanocyte-specific gene / melanoma / melanin synthesis / siRNA発現HIVベクター / 色素細胞発現遺伝子 / siRNAオリゴヌクレオチド / 色素細胞特異遺伝子 |
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| Research Abstract |
We attempted to analyze function of melanocyte-specific genes(MART-1 and AIM-1) and highly expressed genes (β-catenin and FABP7) in melanoma by suppression of gene expression using RNA interference. First we constructed a measurement system that could deduce efficiency of RNA interference by the content of melanin in pigmented melanoma cells. Then we tried to downregulate the MART-1 and the AIM-1 expression with each specific siRNA. The expression of MART-1 and AIM-1 was lowered to 50 and 30 %, respectively, of that in control cells. But there were no changes in the phenotype of melanoma cells at least in growth and migration. β-catenin known as an aberrant accumulated protein in melanoma cells was downregulated with β-catenin specific siRNA. β-catenin protein of 624Amel and 888mel with specific siRNA was decreased less than 20% of control cells and cell proliferation(WST-1 assay) of 624Amel and 888mel with specific siRNA was decreased to 70-80% compared to normal cells. FABP7 identified as a highly expressed gene in melanoma cell lines by DNA chips was then used. When the FABP7 expression was downregulated with FABP7 specific siRNA, in vitro cell proliferation and Matrigel migration of melanoma cell lines, WM266mel and 888mel, were decreased. In contrary, cell proliferation and Matrigel migration of 293T cells with plasmid FABP7 was increased. These results suggest that FABP7 may be involved in formation of malignant phenotype of melanoma. We could not detect deference of melanin content or other phenotypes between RNA interfered cells and control cells about MART-1 and AIM-1. In melanoma cells with β-catenin and FABP7 specific siRNA, we could observe suppression of cell proliferation, and we can also detect suppression of cell migration about FABP7. We think RNA interference is a very useful tool for development of functional analysis and molecular target therapy.
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