C-MYC antisense oligodeoxynucleotides can induce apoptosis and down-regulate Fas expression in rheumatoid synoviocytes

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Abstract
Objective To investigate the role of c‐myc in the pathogenesis of rheumatoid arthritis (RA) and the mechanism of synovial apoptosis. Methods Using cultured human synoviocytes from patients with RA and c‐myc antisense oligodeoxynucleotides (AS ODN), we examined the inhibition of cell proliferation by the MTT assay and the induction of apoptosis with TUNEL staining and fluorescence microscopy. In addition, the effect of c‐myc on down‐regulation of Fas expression was analyzed by flow cytometry, cytotoxicity assay, and reverse transcriptase–polymerase chain reaction. Results Treatment with c‐myc AS ODN induced inhibition of cell proliferation, along with down‐regulation of c‐Myc protein and c‐myc messenger RNA (mRNA) expression. The morphologic changes of synovial cell death were typical of apoptosis. In addition, c‐myc AS ODN treatment down‐regulated expression of Fas mRNA but not Fas antigen. Analysis of the involvement of the caspase cascade revealed that the cytotoxic activity of c‐myc AS ODN was completely blocked by inhibitors of both caspase 1 (YVAD–FMK) and caspase 3 (DEVD‐FMK). Conclusion Our results strongly suggest that c‐myc AS ODN might be a useful therapeutic tool in RA and clarify that cell death by c‐myc AS ODN is induced through the caspase cascade, similar to Fas‐induced apoptosis. In addition, combination therapy with anti‐Fas antibody and c‐myc AS ODN reduced Fas‐dependent cytotoxicity.