| Project/Area Number |
09557120
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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 | 展開研究 |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | KUMAMOTO UNIVERSITY |
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Principal Investigator |
USHIO Yukitaka KUMAMOTO UNIV.MED.SCH., PROFESSOR, 医学部, 教授 (20028583)
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| Co-Investigator(Kenkyū-buntansha) |
NISHI Tohru KUMAMOTO UNIV.HOSP., INSTRACTOR, 医学部・附属病院, 助手 (00264309)
KOCHI Masato KUMAMOTO UNIV.MED.SCH., ASSOCIATE PROFESSOR, 医学部, 助教授 (70178218)
KURATSU Jun-ichi KAGOSHIMA UNIV.MED.SCH., PROFESSOR, 医学部, 教授 (20145296)
NAKANO Masahiro KUMAMOTO UNIV.HOSP., PROFESSOR, 医学部・附属病院, 教授 (40002125)
SAYA Hideyuki KUMAMOTO UNIV.MED.SCH., PROFESSOR, 医学部, 教授 (80264282)
竹島 秀雄 熊本大学, 医学部, 助手 (70244134)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥8,200,000 (Direct Cost: ¥8,200,000)
Fiscal Year 1998: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1997: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | Invasin / microsphere / anti-cancer drug / GST-fusion protein |
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| Research Abstract |
1. Bsic research on the biological property of invasin protein We have developed a method for mass production of the GST-invasin fusion protein by using recombinant technique. This protein has been proved to have biological activity of invasin. This notion was consistent with the fact that the C-terminal of invasin protein is essential for its binding to a receptor. I has been reported that only a single domain consisted of 192 residues at the C-terminal end of the invasin protein is required for the binding and inasion to cells. We produced several deletion mutant proteins of invsin, and identified the minimum sequence in the invasin protein for the invaion into mammalian cells. This partial protein was produced as a GST-fusion protein in B.coli and purified by using a GSH-beads column. This purified protein was added into culture medium of human fibroblast and several kinds of tumor cell line as a monomar or polymar, and invasion of those proteins were observed. It was proved that th
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e assembly of integrin b is essential in this steps. Furthermore, it was proved that invsin induced phagocytosis was suppressed by actin polymerization while it was promoted by microtubules assembly by the experiments using several drugs which affect cytoskeleton reorganization.
2. Developement of a microsphere which can be conjugated with invasin protein
Invasin protein can not induce phagocytosis as a monomar protein. It needs to assemble to have biological activity. Therefore, simple conjugation of invasin protein and anti-cancer drug can not achieve effeicent delivery of the drug into cancer cells. We planned to deliver thoes drugs into/around cancer cells by conjugating GST-fusion invsin and microsphere containing anti-cancer drugs. This drug can be substituted by some kinds of genes. Polydepsipeptide was suitablematerial to make microsphere for the purpose of cunjugation of GST-fusion protein. As a basic test for making microsphere containing less water-soluble anti-cancer drug, we succeeded to construct a microsphere made by poly lactic acid. A high trapping efficacy and slow release was obtained by the method. We are now trying to conjugate the fusion protein and microsphere. Less
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