| Project/Area Number |
11680825
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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 |
Laboratory animal science
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| Research Institution | Tokai University |
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Principal Investigator |
UEYAMA Yoshito Tokai University Schcol of Medicine Associate Professorr, 医学部, 助教授 (30072408)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Masato Tokai University School of Medicine Associate Professor, 医学部, 助教授 (00164335)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | ribozyme / vascular endothelial growth factor / pancreatic cancer / isoform / distant metastasis / cancer gene therapy model / angiogenesis / 肝転移モデル / 免疫不全動物 |
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| Research Abstract |
Stromal angiogenesis is an important factor for progression of malignant neoplasms. we used hammerhead ribozymes against vascular endothelial growth factor (VEGF) gene transcripts to downregulate cell-associated VEGF189 isoform function in a pancreatic cancer cell line MIA-PaCa2. The b-actin promoter-driven ribozyme sequences were introduced into the cells by lipofection. We evaluated their effects on growth and neovascularisation of the cell lines. Metastatic potential was evaluated by the injection of transformants into the splenic pulp of nude mice. Anti-VEGF189 ribozyme specifically and efficiently cleaved the VEGF189 mRNA isoform but not other types of isoforms in vitro. MIA-Paca2 introduced with anti-VEGF189 ribozyme did not show any alteration of growth rate under tissue culture conditions compared to MIA-Paca2 introduced with disabled ribozyme. Production of tissue type plasminogen activator decreased in the human umblical vascular endothelial cells cocultured with the MIA-Paca2 cell line introduced with anti-VEGF189. When the transformants were subcutaneously transplanted, tumour volume of the ribozyme-introduced MIA-Paca2 xenografts was significantly smaller than that of disabled ribozyme-introduced cell line xenografts (P<0.01, Dunnett's test). No metastasis was apparent of the MIA-Paca2 introduced with anti-VEGF189, while disabled ribozyme transfected MIA-Paca2 show significant liver metastasis (P<0.05, Fisher's exact test). These results suggested that the VEGF189 mRNA plays an important role in growth and metastatic potential through alteration of angiogenic balance in cancer tissue.
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