| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2002: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Research Abstract |
[Purpose]
We investigated the role of heparin binding domein of fibronectin expressing around carcinoma cells, using several carcinoma cell lines,mainly,gastric cell line (STKM) and sodium heparin.
[Materials and Methods]
1) Immunocytochemistry and flowcytometry has been done to STKM cells to confirm the expression of FN.
2) Sodium heparin was added at the concentration of 10, 50,100,200,300 IU/ml to STKM cells. The cells with heparin were investigated cell adhesion by colonmetnc assay,and cell death by thyimdine incorporated assay.
3) The monoclonal antibody against c-terminal heparin binding site of FN, IST-land IST-2 were added to STKM cells and assayed by colorimetnc, and thymidin incorporate methods.
4) bcl-2, and bcl-xl gene were transfected to STKM cells by lipofectin method, and added heparin.The transfected cells with heparin were also assayed by colorimetnc, and thymidin incorporate methods.
[Results]
1 Heparin induces apoptosis via heparin binding sites of FN, which is expressed around carcilnoma cells
2 The results of experiment, using monoclonal antibodies against heparin binding site suggest that the apoptotic effects by heparin is not related to anticoagulant effect of heparin.
3 The apoptosis induced heparin causes via mitochondria inner cells.
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