| Project/Area Number |
13671942
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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 |
Functional basic dentistry
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| Research Institution | Saga Medical School |
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Principal Investigator |
KUKITA Akiko Saga Medical School, Medicine, Associate Professor, 医学部, 助教授 (30153266)
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| Co-Investigator(Kenkyū-buntansha) |
KUKITA Toshio Saga Medical School, Dentistry, Associate Professor, 大学院・歯学研究院, 助教授 (70150464)
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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 |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | osteoclast / differentiation / macrophage / histone deacetylate inhibitor / 転写因子 |
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| Research Abstract |
(1) Analysis of role of OCZF in the signal transduction of osteoclastogenesis
OCZF inhibit transcriptional activity of macrophage transcriptional factor, egr-1. Histone deacetylase inhibitor, trichostatin A (TSA) inhibits transcriptional repressive activity of OCZF. TSA and a another histone deacerylase inhibitor, sodium butyrate, NaB inhibited osteoclast differentiation. On the other hand, TSA and NaB did not have the effect on the differentiation into macrophages. In a macrophage cell line, RAW 264 which is able to differentiate into osteoclasts by the addition of RANKL, TSA and NaB inhibited the translocation of NFkappaB into nucleus and phospholylation of MAPK, whose are regulators of osteoclastogenesis. Data suggest the target genes of T5A and NaB may be regulated by OCZF and involved in the signal transduction of osteoclastogenesis.
Stimulative activity of NFkB of OCZF was analyzed. OCZF did not affect the binding activity to this consensus sequence. On the other hand, we found OCZ
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F stimulated the transcription from the reporter plasmids which have promoter region of p21 and p27, cyclin-dependent kinase (CDK) inhibitors whose expression is stimulated by ODF in osteoclastogenesis. Difference of the expression of protein and mRNA between macrophage cell, RAW264 expressing OCZF and control was compared by western analysis and DNA microarray.
(2) Analysis of OCZF expression by in situ hybridization of rat adjuvant arthritis.
In situ hybridization analysis revealed that OCZF is strongly expressed in osteoclasts and some cells in germinal center.
(3) Analysis of signaling molecules interacting with OCZF
We isolated RNA and prepared cDNA expressed in RAW 264 cells and are analyzing some genes which are possibly interacting with OCZF with yeast two-hybrid system. At same time we establish the cell line in which the expression of OCZF is regulated by tetracycline. Using the cell lines, we are going to find out the molecules which interact with OCZF by immunoprecipitation methods. Less
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