Genistein Inhibit Cytokines or Growth Factor-Induced Proliferation and Transformation Phenotype in Fibroblast-Like Synoviocytes of Rheumatoid Arthritis

The purpose of this research is to study the effect of genistein on cytokines or growth factor-induced proliferation and transformation phenotype of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). RA-FLS were primarily cultured. With respective stimulation of IL-1 beta, TNF-alpha, and EGF, genistein was applied to elucidate its effect on synoviocytes' growth number, cell proliferation assay, cell cycle using cell counts, H-3-TdR incorporation and flow cytometry, the colony numbers under anchorage-independent condition, and the expression of MMP-2 and MMP-9 in synovial fibroblasts. EGF, IL-1 beta, and TNF-alpha increased H-3 incorporation in RA-FLS, respectively. EGF augmented clone numbers of RA-FLS under anchorage-independent condition and not IL-1 beta and TNF-alpha. Genistein had an inhibitory role on cell number and H-3-TdR incorporation of RA-FLS stimulated with IL-1 beta, TNF-alpha and EGF; genistein arrested the cell cycle at G(1) restriction point; genistein decreased colony numbers under anchorage-independent condition stimulated by EGF in serum condition. IL-1 beta or TNF-alpha increased expression of MMP-9 and MMP-2 in rheumatoid synoviocytes; EGF stimulated expression of MMP-9 but not of MMP-2; genistein suppressed production of MMP-9 more than MMP-2 induced by IL-1 beta or TNF-alpha; rMMP-9, rMMP-2, or their inhibitors had no effect on the H-3-TdR incorporation of synovial cells. Erk1/2 inhibitor (PD098 059) had obvious inhibitory effect on the H-3 incorporation induced by TNF-alpha or IL-1 beta; inhibitors of JNK (SP600 125) had no significant effect on the H-3 incorporation. While pretreatment with PD098059 had no marked inhibitory effect on MMP-9 expression induced by TNF-alpha or IL-1 beta, SP600125 decreased significantly the MMP-9 expression induced by TNF-alpha or IL-1 beta. Neither PD098059 nor SP600 125 could inhibit the MMP-2 expression induced by TNF-alpha or IL-1 beta. Genistein inhibited IL-1 beta, TNF-alpha or EGF-induced proliferation and MMP-9 expression in fibroblast-like synoviocytes of rheumatoid arthritis; the proliferation of RA-FLS was mediated by Erk1/2 but not JNK activation, while JNK activation was involved in the signal transduction pathway leading to MMP-9 expression in rheumatoid synoviocytes.