| Project/Area Number |
17591449
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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 |
Digestive surgery
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| Research Institution | Hyogo College of Medicine |
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Principal Investigator |
HIRANO Tadamichi Hyogo College of Medicine, Faculty of Medicine, Research Associate, 医学部, 助手 (90340968)
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| Co-Investigator(Kenkyū-buntansha) |
FUJIMOTO Jiro Hyogo College of Medicine, Faculty of Medicine, Professor, 医学部, 教授 (90199373)
IIMURO Yuji Hyogo College of Medicine, Faculty of Medicine, Associate Professor, 医学部, 助教授 (30252018)
SUGIMOTO Takaaki Hyogo College of Medicine, Faculty of Medicine, Research Associate, 医学部, 助手 (60399152)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | NK4 / angiogenesis of tumor / Gene Therapy / Hepatocellular carcinoma / HGF / 肝細胞癌 / 骨髄移植 / 遺伝子導入 |
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| Research Abstract |
As a new strategy for hepatocellular carcinoma, we have reported NK4 mediated gene therapy that could inhibit tumor angiogenesis, metastasis, and invasion. It markedly suppressed tumor growth and invasion not only in vitro but also in vivo. Based on this study, we aimed to elucidate the mechanism of NK4 mediated anti-angiogenesis, and estimated method of gene transfer for applying human clinical trial. (1) Although NK4 inhibits growth and migration of small-vessel endothelial cells, an inhibitory effect of NK4 in BM-derived cells has not been shown. We attempted to assess whether NK4 suppressed recruitment of BM-derived endothelial cells in tumor vessels using lethally irradiated, tumor-bearing athymic nude mice transplanted with LacZ^+ bone marrow (BM) cells. Nude mice were lethally irradiated (4.5 Gray, with cesium-137 as the source). LacZ^+ bone marrow (BM) cells (5x10^6) obtained from transgenic mice were injected into tail veins of irradiated nude mice. After 4 weeks, these mice r
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eceived injections in the flank of 10^7 HUH7 cells. At 14 days post tumor cell implantation ( or when tumor volume reached approximately 500 mm3 ), either Ad.NK4 (10^9 pfu) or PBS (50 μl) was injected directly into the growing tumor. After 5 days the tumor was removed and stained with anti-lacZ rabbit polyclonal antibody and anti-CD31 rat monoclonal antibody. In all mice most of the original splenic cell population was replaced by donor cells. However, no cells were positive for LacZ by immunohistochemical staining in CD31-positive tumor vessels, even in PBS-injected control mice. In NK4-treated tumors, CD31-positive vessels were significantly fewer than in LacZ-treated controls. These results indicate that NK4 suppressed tumor angiogenesis, although any s uppressive effect for BM-derived e ndothelial cells remains unknown in the present animal model. Further study of this issue is required, since it has important implications for future approaches to of anti-angiogenic therapy. (2) To assess the NK4 influence for liver regeneration, Ad.Nk4 was infected t o mice t hat were performed 70 % hepatectomy. Liver regeneration significantly delayed regarding to control that were infected Ad.LacZ, as a result of hepatic weight, PCNA staining and BrDU. (3) Efficacy of hydrodynamics transfection mediated NK4 transduction was assessed instead of adenoviral vector to aid human clinical trial. Suppression of tumor growth was confirmed by this transfection method. Less
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