| Project/Area Number |
14571350
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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 |
Cerebral neurosurgery
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| Research Institution | Chiba Cancer Center Research Institute |
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Principal Investigator |
OGA Masaru Chiba Cancer Center Research Institute, Division of Neurological Surgery/Division of Chemotherapy, 脳神経外科・化学療法研究部, 医長・兼務研究員 (10251159)
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| Co-Investigator(Kenkyū-buntansha) |
HATANO Kazuo Chiba Cancer Center Research Institute, Division of Radiation Oncology, Head, 放射線治療部, 部長
TAKENAGA Keizo Chiba Cancer Center Research Institute, Division of Chemotherapy, Chief Scientist, 化学療法研究部, 主席研究員 (80260256)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Radio-sensitization / Egr-1 / CArG element / gene therapy / Glioma / Suicide gene |
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| Research Abstract |
In the first year we verified that the gene expression located at the downstream of radio-sensitized promoter would actually amplify by radiation. We constructed the EGFP&HSV-tk vectors containing E_4/CMV chimeric radio-sensitized promoter (E_4: four-repeated structure of CArG element located at Egr-1 gene). E_4/EGFP was transferred into U251-MG/U373-MG human glioma cell line respectively and the expression of EGFP was evaluated by optical observation under fluorescent microscopy and Western blotting after 3Gy irradiation. As a result, we verified that the expression of EGFP was amplified about 1.3〜1.4 times after 6〜54 hours compared to pre-irradiation stage. Then E4/HSV-tk and HSV-tk were transferred into U251-MG/U373-MG human glioma cell line respectively, and the degree of cell proliferation was evaluated by MTT assay after irradiation (1,3,5Gy) following GCV administration. The results were 1)the radio-sensitized promoter also could be activated by physical/chemical stimuli. 2)the radiation dose-dependent apoptosis was observed only in the E_4/HSV-tk-administrative group but not statistically significant.
In the second year, we replaced the promoter with E_<9ns-2>, stronger than E_4 in radio-sensitization and did the same trial. Contrary to the expectation, however, the expression of EGFP was not significantly amplified in the E_<9ns-2>/EGFP -administrative groups. In the MTT assay, we also could not verified the significant inhibition of the cell proliferation among E_<9ns-2>/HSV-tk-administrative glioma cell lines after irradiation. We are going to promote our previously scheduled plan by using E_4/CMV as the radio-sensitized promoter, transferring E_4/HSV-tk into C6 and endothelial precursor cell line.
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