| Project/Area Number |
08045062
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | University-to-University Cooperative Research |
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| Research Field |
General medical chemistry
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| Research Institution | TOKYO INSTITUTE OF TECHNOLOGY |
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Principal Investigator |
SAITO Yuji (Tokyo Institute of Technology, Faculty of Bioscience and Biotechnology, Associate Professor), 生命理工学部, 助教授 (30134810)
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| Co-Investigator(Kenkyū-buntansha) |
BLEACKLEY Christopher ( The University of Alberta, Department of Biochemistry, Professor), Department of, 教授
MCELHANEY Ro The University of Alberta, Department of, 教授
TAKAGI Junichi (Tokyo Institute of Technology, Faculty of Bioscience and Biotechnology, Assist, 生命理工学部, 助手 (90212000)
MCELHANEY Ronald N. (The University of Alberta, Department of Biochemistry, Professor)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | tumor cells / platelet aggregation / blood coagulation / Factor X / Factor V / thrombin / monoclonal antibody / cloning / モノクローナル抗体 / 血小板凝固 / モノクロナール抗体 / サブトラクションクローニング / 組織因子 / EPR-1 / 分化誘導 |
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| Research Abstract |
One of the main causes of death of cancer patients is thrombosis. This is probably because cancer cells activate platelet aggregation and blood coagulation. Furthermore, it is fairly well accepted that above phenomena might be responsible for metastasis of cancer. In the initial year of our research program we did following experiments. We found using a neuroblastoma cell GOTO that platelet aggregation was induced upon addition of this cell to platelet-rich plasma. We also found that the addition of GOTO cells to normal plasma activated Factor X and it in turn activated prothrombin and produced thrombin. We then found that both activities, the activation of Factor X and prothrombin, disappeared upon differentiation of GOTO cells to Shwanian-like cells. Based on the results obtained in the initial year, we did subtraction cloning and production of monoclonal antibodies in the second year with the aim of elucidating the underlining mechanism of thrombin production by GOTO cells in molecular terms. In the final year we are looking into the specific binding of Factor X to the cells to carry out an expression cloning. As far as the production of monoclonal antibodies is concerned, we have been able to clone hybridomas directly on soft : agar plates and we have obtained several clones with the desired properties. We hope the time will soon come to start the actual cloning experiment with these antibodies. On the other hand we have made much improvement as to the purification of highly polar lipid, probably glycolipid, which is the antigen for the monoclonal antibody we established last year which specifically recognized the surface of neuroblastoma cells. We hope we could do NMR and mass spectrometric analyses to elucidate the chemical structure of this interesting lipid. We will chemically synthesize the lipid and elucidate the biological function of this lipid.
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