| Project/Area Number |
11671366
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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 | KYOTO UNIVERSITY |
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Principal Investigator |
TAKAHASHI Jun Kyoto University, Graduate School of Medicine, Lecturer, 医学研究科, 講師 (80252435)
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| Co-Investigator(Kenkyū-buntansha) |
UEBA Tetsuya Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (00314203)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | glioma / FGF-2 / transcription / tumor suppressor gene / Transcription factor / brain tumor |
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| Research Abstract |
Object : Fibroblast growth factor-2 (FGF-2) is know to play an important role inangiogenesis and in tumor growth of astrocytomas. The goal of the study is theelucidation of a mechanism that controls FGF-2 gene expression.
Results : In search of the mechanism that regulates FGF-2 expression, we have focused on a tandem repeat sequence (gccgaac) around the transcription initiation site of the FGF-2 promoter. Using this sequence as bait, we conducted yeast-one-hybrid screening and cloned a 24kDa protein (p24) that could bind the promoter region of FGF-2. To confirm the affinity of p24 to the gccgaac sequence, we performed gel mobility shift assay. P24 shows highly specific affinity to gccgaac sequence. Reverse transcriptase (RT)-PCR showed that p24is expressed neither in U87MG nor in U251MG glioma cell lines. Immunocytochemistry shows that p24 expressing under the control oftetracycline-responsive promoter (tetracyclme-on system) is localized mainly in the nucleus of U87MG glioma cells. Luciferase assay revealed that p24 repressed the FGF-2 promoter activity about 10-fold in the U87MG cell line and about five fold in the U251MG cell line (p<0.05). Quantitive RT-PCR using Light Cycler (Roche, Go. Ltd.) also revealed that p24 repressed the endogenous expression of FGF-2 gene about ten fold (p=0.0010).
Conclusion : Our data show that p24 represses the gene expression ofFGF-2 in glioma cells as a transcription factor. Further investigation of the significance of p24 in vivo is going on using surgical specimens.
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