| Project/Area Number |
13671555
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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 |
Orthopaedic surgery
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| Research Institution | Chiba Cancer Center Research Institute |
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Principal Investigator |
NAKAMURA Yohko Chiba Cancer Center Research Institute, Division of Biochemistry, Research Fellow, 生化学研究部, 研究員 (60260254)
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| Co-Investigator(Kenkyū-buntansha) |
ISOGAI Eriko Chiba Cancer Center Research Institute Division of Biochemistry Research Fellow, 生化学研究部, 研究員 (40300917)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | BMP / neuronal differentiation / neuroblastoma / DAN / p53 family / P27^<KIP1> / osteoblast |
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| Research Abstract |
DAN, a putative tumor suppressor, was reported to antagonize BMP in Xenopus Oocyts. We investigated the interaction of DAN with BMP2 during osteoblast differentiation and also in transformed cells. A possibility that DAN could bind BMP2 through the cysteine- rich domain was confirmed by in vitro binding assay. During osteoblastic differentiation of mouse osteoblast MC3T3-E1 cells which were treated with ascorbic acid for 6 days, the activity of alkaline phosphatase (ALP)an osteoblastic differentiation marker, was up-regulated while the level of DAN was increased. The activity of ALP in osteoblasts of primary culture prepared from the DAN knockout mouse was down-regulated compared with wild type mouse. Therefore, it was suggested that DAN is regulated during osteoblast differentiation through the interaction with BMP2.
To examine whether human neuroblastoma-derived cell lines could respond to BMP treatment, we analyzed phosphorylation of Smad1/5 in the presence of BMP2. Phosphorylated fo
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rm of Smad1/5 showed a remarkable increase at 30 min after the addition of BMP2. These observations strongly suggest that BMP signaling cascade exists in human neuroblastoma-derived cell lines. The BMP2-dependent neurite outgrowth was also detectable accompanied with the growth inhibition. We have recently found that p73 , a member of p53 family, transactivates the transcription of DAN. To investigate whether functional interaction between DAN and p53 family members could also be involved in the regulation of the BMP-induced differentiation of neuroblastoma cells, we examined the expression levels of DAN and p53 family members in response to BMP2. BMP2 treatment resulted in a significant down-regulation of DAN, p73 and p53 expression in a time-dependent manner. Furthermore, accumulation of Cdk inhibitor, p27^<KIP1> and reduction of Cdk2 activity were detected in BMP2-treated cells. These results suggest that p27^<KIP1> plays a crucial role in the BMP2- induced growth arrest and neuronal differentiation of neuroblastoma cells. Less
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