| Project/Area Number |
06671691
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | The Jikei University School of Medicine |
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Principal Investigator |
SASAKI Hiroshi The Jikei University School of Medicine, LECTURER, 医学部, 講師 (80125020)
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| Co-Investigator(Kenkyū-buntansha) |
TADA Akio The Jikei University School of Medicine, LECTURER, 医学部, 助手 (40197364)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1995: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Endometrial Neoplasms / Estrone Sulfatase / Monoclonal antibody / Immunohistochemistry / 子宮内膜癌 / 遺伝子移入 |
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| Research Abstract |
Endometrial carcinoma tissues showed locally high activity of estrogen based on high activity of estrtone-sulfatase. However, histrological localization of the enzyme has not been unknown, because anti-cstrone-sulfatase monoclonal antibody is not available yet. The purpose of present study is the production of the monoclonal antibody and in order to demonstrate the High-activity of estrone-sulfatase in endometrial carcinoma tissues has been reported.histological localization of estrone sulfatase in endomentrial tidsses. Ten placentas were homogenized and cluted by columchromatography. Enzyme assay with H3-estrone-sulfate showed a high rich peak of estrone-sulfatase. The protein on the peak were eluted and was used as an antigen for the production of monoclonal antibody. BALB/c nu/nu mice were immunized with the protein and then twelve clone cells were selected by both ELISA and western-blotting. Immunohistochemistry showed that the all monoclonal antibodies were reactive to normal placenta, but inactive to placentas of congenital ichtyosis. Transfection with aryl-sufatase-C gene to breast carcinoma cell lines (MCF-7 and T47D) showed higher activities of both estrone-sulfatse and dehydroepiandrosterone than transfectants with vector alone. The monoclonal antibodies reacted on the enzyme proteins from the transfectants. This proves that the monoclonal antibodies were detected both estrone-sulfatase and dehydroepiandrosterone, Immunohistochemical staining of endometrial tissues and neoplasms demonstrated that normal endometrial tissues were inactive, but endometrial acrcinoma tissues, especially carcinoma cells alone, were reactive to the antibody. This suggests that estrone-sulfatase are localized on epithelial cells of endometrial carcinomas. In conclusion, we succeeded in the production of anti-estrone-sulfatase monoclonal antibodies.
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