Trichostatin A sensitises rheumatoid arthritis synovial fibroblasts for TRAIL-induced apoptosis

• Published in: Annals of the rheumatic diseases 2006-07, Vol.65 (7), p.910-912
• Main Authors: Jüngel, A, Baresova, V, Ospelt, C, Simmen, B R, Michel, B A, Gay, R E, Gay, S, Seemayer, C A, Neidhart, M
• Format: Article
• Language: English
• Subjects: Aged; Apoptosis; Apoptosis - drug effects; Apoptosis Regulatory Proteins - pharmacology; Arthritis, Rheumatoid - metabolism; Arthritis, Rheumatoid - pathology; Biological and medical sciences; Blotting, Western - methods; Cancer; Cell cycle; Cell growth; Cells, Cultured; Cyclin-Dependent Kinase Inhibitor p21 - analysis; Diseases of the osteoarticular system; Dose-Response Relationship, Drug; Drug Synergism; Extended Report; Female; Fibroblasts; Fibroblasts - drug effects; Fibroblasts - metabolism; Fibroblasts - pathology; Flow Cytometry; Gangrene; Gene expression; HDAC; histone acetylation/deacetylation; Humans; Hydroxamic Acids - pharmacology; Immunoglobulins; Inflammatory joint diseases; Male; Medical sciences; Membrane Glycoproteins - pharmacology; Middle Aged; osteoarthritis; Osteoarthritis - metabolism; Osteoarthritis - pathology; RASF; Rheumatoid arthritis; rheumatoid arthritis synovial fibroblasts; Streptomyces; Studies; synovial fibroblasts; Synovial Membrane - drug effects; Synovial Membrane - metabolism; Synovial Membrane - pathology; TBST; TNF-Related Apoptosis-Inducing Ligand; TRAIL; trichostatin A; Tris buffered saline containing 0.1% Tween 20; TSA; Tumor Necrosis Factor-alpha - pharmacology; Tumor necrosis factor-TNF; tumour necrosis factor related apoptosis-inducing ligand
• ISSN: 0003-4967; 1468-2060
• DOI: 10.1136/ard.2005.044065

Background: Histone acetylation/deacetylation has a critical role in the regulation of transcription by altering the chromatin structure. Objective: To analyse the effect of trichostatin A (TSA), a streptomyces metabolite which specifically inhibits mammalian histone deacetylases, on TRAIL-induced apoptosis of rheumatoid arthritis synovial fibroblasts (RASF). Methods: Apoptotic cells were detected after co-treatment of RASF with TRAIL (200 ng/ml) and TSA (0.5, 1, and 2 μmol/l) by flow cytometry using propidium iodide/annexin-V-FITC staining. Cell proliferation was assessed using the MTS proliferation test. Induction of the cell cycle inhibitor p21Waf/Cip1 by TSA was analysed by western blot. Expression of the TRAIL receptor-2 (DR5) on the cell surface of RASF was analysed by flow cytometry. Levels of soluble TRAIL were measured in synovial fluid of patients with RA and osteoarthritis (OA) by ELISA. Results: Co-treatment of the cells with TSA and TRAIL induced cell death in a synergistic and dose dependent manner, whereas TRAIL and TSA alone had no effect or only a modest effect. RASF express DR5 (TRAIL receptor 2), but treatment of the cells with TSA for 24 hours did not change the expression level of DR5, as it is shown for cancer cells. TSA induced cell cycle arrest in RASF through up regulation of p21Waf1/Cip1. Levels of soluble TRAIL were significantly higher in RA than in OA synovial fluids. Conclusion: Because TSA sensitises RASF for TRAIL-induced apoptosis, it is concluded that TSA discloses sensitive sites in the cascade of TRAIL signalling and may represent a new principle for the treatment of RA.