2005 Fiscal Year Final Research Report Summary
Roles for histone deacetylases in apoptosis and proliferation of RA synovial cells : implications for therapy
| Project/Area Number |
16590981
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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 |
膠原病・アレルギー・感染症内科学
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| Research Institution | Kobe University |
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Principal Investigator |
MORINOBU Akio Kobe University, Graduate School of Medicine, Lecturer, 大学院・医学系研究科, 講師 (10294216)
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| Co-Investigator(Kenkyū-buntansha) |
KUMAGAI Shunichi Kobe University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (00153346)
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| Project Period (FY) |
2004 – 2005
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| Keywords | Rheumatoid Arthritis / synovial cells / HDAC inhibitor / Fas signaling / Apoptosis |
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| Research Abstract |
1.Effects of HDAC inhibitors on cultured RA-synovial cells
Objective : In order to clarify the effects of trichostatin A (TSA), a histone deacetylase inhibitor, on the growth and survival of rheumatoid arthritis synovial fibroblasts (RA-SF). Methods : Cell viability was assessed using a WST-8 assay and direct cell counting. Apoptosis was detected by annexin V staining on a flowcytometer. Protein and mRNA expression was determined by western blotting, flow cytmetry, and RT-PCR. Results : TSA suppressed cell growth of RA-SF in a dose dependent manner, determined by WST-8 assay and direct cell counting. Other histone deacetylase inhibitors also showed inhibitory effects on RA-SF proliferation. TSA up-regulated p21^<WAF1/CIP1> cell cycle inhibitor, suggesting that cell cycle arrest is involved in the reduction of cell number. In addition, TSA co-operated with Fas-induced pathway to induce cell death, determined by WST-8 assay and annexin V staining. TSA reduced FLIP expression but not Bcl-2, Bcl-X_L, and Fas expression, indicating that the synergistic effect may be through down-regulation of FLIP. Conclusion : TSA has anti-rheumatic effects on RA-SF and might be a potential therapeutic tool for the treatment of RA.
2.Genes that are regulated by TSA, an HDACi, in RA-SF
In order to determine which genes are up- or down- regulated by TSA in RASF, cDNA subtraction and differential display technique were carried out. Differentially regulated genes were cloned, sequenced and identified by Blastn program. Some of the candidate genes were subjected to quantitative RT-PCR to confirm that they are regulated by TSA. At least 2 genes were determined to be regulated by TSA.
3.Animal study were performed using Ab-induced arthritis model.
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