| Project/Area Number |
12670147
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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 |
Pathological medical chemistry
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| Research Institution | Tokai University |
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Principal Investigator |
TAMALA Tomoo Tokai University, Tokai University, Scool of Medicine, Assistant Researcher (50192175)
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| Co-Investigator(Kenkyū-buntansha) |
KIJIMA Hiroshi Tokai University, School of Medicine, Assistant Researcher (90204859)
TANAKA Manami National Institute of Advanced Industrial Science and Technology, Department of Collaborations, Head of Bradeion Project
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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 |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2001: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Bradeion / colorectal cancer / septin family / gene diagnosis / gene therapy / chimera mouse / structual analysis / テストストリップ / 構造解析 |
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| Research Abstract |
Bradeion, for colorectal cancer, was discovered to be a septin family gene related to cell division and growth control by our research group. The Bradeion gene can be considered as a marker for colorectal cancer in the following reasons:
(1) selectivity of cell types; (2) specificity ("all or none" type expression changes); (3) neither any amino acid nor structural changes caused by point mutations, deletions, miss-sense mutation, etc. in the course of phenotype change.
We carried out the basic study of a (gene) diagnostic method and application to a gene therapy method using Bradeion for the colorectal cancer.
1. Development of a simple diagnostic method
(1) Genetic screening method
Bradeion RT-PCR primers were designed and synthesized, and RT-PCR using cultivated cancer cells and patient tissues. As the results of these studies, it became clear that a Bradeion RT-PCR diagnostic method is useful.
(2) Application for test strip method
As Bradeion fusion protein was expressed, extracted and purified, specific monoclonal antibodies were made. They were evaluated by ELISA, immunohistochemistry and Western blot, and found useful. Further studies are proceeded for clinical application to a test strip.
2. Physiological function analysis of Bradeion
(1) We succeeded in making of a chimera mouse (during a patent application).
(2) We succeeded in introduction of Bradeion gene to fruit fly (Jap. Patent 2001-54759) to discover Bradeion -relating materials by Two-hybrid method.
3. Development of the gene therapy by a control of Bradeion expression.
(1) We discovered that G2 arrest in a cell cycle was occurred by Bradeion-specific ribozymes introduction to a cultivated colorectal cancer cells.
(2) To develop gene therapy method of colorectal cancer, colorectal cancer cells in which Bradeion-specific ribozymes were transfected were inoculated in nude mice. Then the masses of colorectal cancer cells with Bradeion -specific ribozymes did not grow and smaller than those of negative control mice.
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