| Project/Area Number |
16580078
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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 |
Applied biochemistry
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| Research Institution | Chubu University |
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Principal Investigator |
OHNISHI Motoko Chubu University, College of Bioscience and Biotechnology, Associate Professor, 応用生物学部, 助教授 (00312653)
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| Co-Investigator(Kenkyū-buntansha) |
WOO JT Chubu University, College of Bioscience and Biotechnology, Professor, 応用生物学部, 教授 (20272693)
NAGAI Kazuo Chubu University, College of Bioscience and Biotechnology, Professor, 応用生物学部, 教授 (00011974)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | signal transduction / development, differentiation / enzyme / biological activity / cells, tissues / 破骨細胞 / プロテインホスファターゼ2C / 破骨細胞分化誘導因子 / RANKL |
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| Research Abstract |
The receptor activator of NF-κB ligand (RANKL), which is a member of the tumor necrosis factor family, plays a key role in the differentiation, activation and survival of the osteoclasts. RANKL interacts with its receptor, RANK, which is expressed on osteoclast progenitors or mature osteoclasts, and then induce activation of downstream molecules such as NF-κB and p38 MAPK. Since protein phosphatase 2C (PP2C) has been reported as one of the negative regulators in SAPK/p38 signaling pathway, we examined the possibility if PP2C might be involved in osteoclastogenesis through the regulation of RANKL/RANK signaling pathway. The following conclusions can be drawn from the results of this research project.
1;When human embryonic kidney epithelial 293 cells were transfected with a full-length RANK(a receptor of RANKL) expression plasmids, phosphorylation level of p38 MAPK and NF-κB activity were upregulated. We found that coexpression of PP2C suppressed activation of both p38 MAPK and NF-κB.
2;We generated 293 cells, which stably express Myc epitope-tagged RANK, called 293-RANK cells. When these cells were stimulated with RANKL, increase in phosphorylation level in p38 MAPK and activation of NF-κB were observed. Overexpression of PP2C inhibited these RANKL-induced p38 phosphorylation and NF-κB activation.
3;RAW264 cells, which are macrophage/monocyte cell line, are known to differentiate into osteoclast-like multinucleated cells following RANKL stimulation. Total mRNAs were prepared from RAW264 cells at regular intervals after RANKL stimulation and quantitative realtime PCR was performed in order to analyze PP2C expression. As a result, it was shown that PP2C mRNA levels increased transiently following RANKL stimulation.
Taken together, our results suggest the possibility that PP2C might be concerned with the regulation of osteoclast differentiation.
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