| Project/Area Number |
09470172
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Kagoshima University |
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Principal Investigator |
KITAJIMA Isao Faculty of Medicine, Kagoshima University, Associate, 医学部, 助教授 (50214797)
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| Co-Investigator(Kenkyū-buntansha) |
UNOKI Kazuhiko Medical Hospital, Kagoshima University, Associate Professor, 医学部・附属病院, 講師 (60193926)
AKASHI Mitsuru Faculty of Engineering, Kagoshima University, Professor, 工学部, 教授 (20145460)
MARUYAMA Ikuro Faculty of Medicine, Kagoshima University, Professor, 医学部, 教授 (20082282)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥12,000,000 (Direct Cost: ¥12,000,000)
Fiscal Year 1999: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1998: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1997: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Angiogenesis / oligodeoxynucleotides / phosphorothioate / bFGF / NF-κB / cornel vascular / Tube formation / antiseise oligodeoxynucleotides / 荷電 / FGF / ホスホロチオエ-ト / ヘパリン / マトリゲル |
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| Research Abstract |
Angiogenesis is regulated by heparin-binding growth factors such as basic fibroblast growth factor. In this study, we investigated the effects of phosphorothioate oligodeoxynucleotides on basic fibroblast growth factor induced angiogenesis in vitro and in vivo. Antisense phosphorothioate oligonucleotides are widely used as specific inhibitors of gene expression. Because phosphorothioate-type oligonucleotides are polyanions, they can also bind many heparin-binding proteins. In an electrophoretic mobility shift assay, phosphorothioate oligonucleotides bound basic fibroblast growth factor in a dose-dependent but sequence-independent manner. However, double-strand phosphothioate oligonucleotide-DNAs did not affect complex formation. On a basement matrix using a Matrigel matrix, we observed less than 50% tube formation by human umbilical endothelial cells with 10 uM basic fibroblast growth factor, vascular endothelial growth factor or Nuclear factor kappa B antisense as well as sense phosph
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orotiate oligodeoxynucleotides, while phosphodiester-type oligodeoxynucleotides were not affected. This effect could be overcome by addition of 1 ug/ml basic fibroblast growth factor. The phosphorothioate but not phosphodiester oligodeoxynucleotides also inhibited the basic fibroblast growth factor-induced rabbit corneal neovascularization. The incubation of basic fibroblast growth factor with nuclear factor-kappa B antisense sequence containing four contiguous guanosine residues significantly abrogated the effects of basic fibroblast growth factor. The nuclear factor-kappa B antisense phosphorotioate oligodeoxynucleotides also showed a low rescue score for basic fibroblast growth factor-dependent photoreceptor rescue due to its degradation by constant light exposer in albino rats. However, antisense phosphorothioate oligodeoxynucleotides active against basic fibroblast growth factor inhibited angiogenesis more strongly than the phosphorothioats oligodeoxynucleotides containing four guanosines. Because of the important role basic fibroblast growth factor plays in angiogenesis, some phosphorothioate oligodeoxynucleotides may serve as potent anti-angiogenic compounds acting through a combination of polyanionic phosphorothioate effects and a sequence-specific antisense mechanism. Less
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