Susceptibility of rheumatoid arthritis synovial fibroblasts to FasL- and TRAIL-induced apoptosis is cell cycle-dependent

Abstract

Introduction The rheumatoid arthritis (RA) synovium is characterised by the presence of an aggressive population of activated synovial fibroblasts (RASFs) that are prominently involved in the destruction of articular cartilage and bone. Accumulating evidence suggests that RASFs are relatively resistant to Fas-ligand (FasL)-induced apoptosis, but the data concerning tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) have been conflicting. Here, we hypothesise that the susceptibility of RASFs to receptor-mediated apoptosis depends on the proliferation status of these cells and therefore analysed the cell cycle dependency of FasL-and TRAIL-induced programmed cell death of RASFs in vitro.

The Effect of Melatonin Adsorbed to Polyethylene Glycol Microspheres on the Survival of MCF-7 Cells

“…Untreated cells were used as negative controls, and cells treated with staurosporine (Sigma [22]), which was used to induce apoptosis, were used as positive controls. The cells were resuspended in 500 μl of binding buffer containing 5 μl of annexin V-FITC (Annexin V-FITC Apoptosis Detection Kit; Alexis Biochemicals, San Diego, Calif., USA) and 5 μl of PI and then incubated for 10 min at room temperature.…”

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“…Untreated cells were used as negative controls, and cells treated with staurosporine (Sigma-Aldrich Co.), an inducer of apoptosis, were used as positive controls. 33 The cells were resuspended in 500 μL of binding buffer containing 5 μL of Annexin V-FITC and 5 μL of propidium iodide (PI) and then incubated for 10 minutes at room temperature. The fluorescence of the cells was analyzed by flow cytometry (FACSCalibur system; BD Biosciences).…”