Role of RICTOR on the tumor necrosis factor-α associated cell activation in rheumatoid arthritis fibrobast-like synoviocytes / 中华风湿病学杂志

ABSTRACT

Objective To study the role of RICTOR on rheumatoid arthritis-fibrobast-like synoviocytes (RA-FLS) activation.Methods FLS were isolated from the primary synovial tissues,which were obtained during joint replacement surgery or arthroscopy from three patients with RA.RA-FLS were stimulated with TNF-α at the dose of 10 ng/ml and 20 ng/ml for 48 h.The expression of RICTOR was detected by western blotting.Chemically synthesized RICTOR gene targeted for double-stranded siRNAs were transfected into RA-FLS by cationic liposome.After being transfected with RICTOR siRNA for 48 h,RA-FLS was treated with or without TNF-α for 48 h.The expression of RICTOR was evaluated by western blotting,and the cell viability was analyzed by methylthiazoltetrazolium (MTT) assay.The data were analyzed using analysis of variance (ANOVA) and LDL-t test.Results The expression of RICTOR protein was significantly higher in the TNF-α stimulated group (at the dose of 10 ng/ml and 20 ng/ml for 48 h) than that in the control group (bothP＜0.05),while the mean change of RICTOR/GAPDH ratios of band optical density x+s was 0.35±0.06 for the control group,0.60±0.09 for the TNF 10 ng/ml group and 1.10±0.12 for the TNF 20 ng/ml group.Moreover,the expression of RICTOR protein was obviously decreased in RICTOR siRNA transfection groupthan that in control after being trans-fected for 96 h (both P＜0.05),and ratios of control group,RICTOR (-)/TNF-α(-) group and RICTOR(-)/TNF-α(+) group was 0.498 4±0.140 1,0.012 8±0.002 0,0.042 5±0.027 3respectively.After the silence of RICTOR,the cell viability decreased in RA-FLS,no matter with or without TNF-α for 48 h later (both P＜0.05).Conclusion These results indicat that RICTOR might play an important role in the TNF-α associated activation of RA-FLS.