| Project/Area Number |
12671970
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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 |
Surgical dentistry
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| Research Institution | Nihon-University |
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Principal Investigator |
IWANARI Shinkichi Nihon-University, School of Dentistry, Research Assistant, 歯学部, 助手 (30168588)
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| Co-Investigator(Kenkyū-buntansha) |
HIROSE Kunitaka Institute of Medical Biology, Kureha chemical Co., Chief investigator, 生物化学研究所・生物探求第2チーム, 課長(研究者)
KOMIYAMA Kazuo Nihon-University, School of Dentistry, Associate Professor, 歯学部, 助教授 (00120452)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | oral cancer / gene therapy / MIP-1α / Mac-1 / tumorigeicity / CCケモカイン / ヌードマウス / 扁平上皮癌 |
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| Research Abstract |
A successful antitumor response by macrophages requires their recruitment to the tumor site along with induction of tumoricidal properties MIP-1α has chemotactic and mitogenic activities toward neutrophils, macrophages, T cell and NK cells, and interacts with the CCR1 and CCR5 receptors.
MIP-1α gene was reconstructed in a palsmid vector and transferred to the HSC-3 cells (HSC-3/MIP-1α) derived from a human oral squamous cell carcinoma, and then transplanted into nude mice to evaluate their tumorigenicity in vivo.
The MIP-1α mRNA expression for HSC-3/MIP-1α was examined by Northern blot analysis. The highest level of MIP-1α mRNA-expressed clone was selected from 10 tested MIP-1α transferred HSC-3 cells and termed as HSC-3/MIP-1αC7. The MIP-1α gene transfection did not affect the cell growth pattern of HSC-3 cells in vitro. The MIP-1α protein was detected by ELISA from HSC-3/ MIP-1αC7 culture supematants as 28.4 ± 0.4 ng/ml.
Both HSC-3/MIP-1αC7 and HSC-3/vector alone developed solid tumors by injecting subcutaneously into nude mice. The tumorigenicity of HSC-3/MIP-1αC7 was clearly reduced in 14th days after the injection compared with HSC-3/vector alone. This reduction was associated with neutrophils and macrophage accumulation that synthesized numerous tumoricidal factors including MIP-1α in the lesion. Local MIP-1αsecretions stimulated those cells, resulting in enhancement of their chemotactic activity. Immunohistopathological staining of HSC-3/MIP-1αC7 injected mice showed numerous infiltration of Mac-1 positive cells into the tumor nest, whereas a small number of infiltrating cells were detected in only HSC-3/vector injected mice. The results indicated that MIP-1α gene transfer to human oral cancer have a potential value of clinical use for the gene therapy.
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