Triad1 Promotes The Apoptosis Of Rheumatoid Arthritis Fibroblast-like Synoviocytes Through P53 Pathway Triad1 Promotes The Apoptosis Of Rheumatoid Arthritis Fibroblast-like Synoviocytes Through P53 Pathway

Objective The purpose of this study is to detect the expression of Triad1 in human rheumatoid arthritis(RA)synovial tissues and explore the possible biological role of Triad1 in the apoptosis of rheumatoid arthritis fibroblast-like synoviocytes(RA FLS).Methods RA synovial tissue samples were collected from 15 patients with RA.Normal synovial tissues were obtained from 6 patients with tibial plateau fracture as control(the 6 patients had no history of RA,and also had no history of meniscus and cruciate ligament injury).The expression of Triad1 was detected in synovial tissues using Western Blot analysis,immunohistochemistry and immunofluorescence.RA FLS isolated from RA patients' synovial tissues were treated with Adriamycin(ADR),a well-known apoptosis-inducing agent,to induce apoptosis.Triad1 protein levels at different time points in the process of ADR-induced apoptosis of RA FLS were detected using Western blot analysis.Expressions and localizations of Triad1 with p53 and C-caspase3 were detected by cell immunofluorescence in RA FLS treated with ADR for 0h and 24 h.After transfection with Triad1-shRNA,the expressions of p53,p21,C-caspase3,Bax and Bcl-2 in RA FLS were detected using Western blot analysis,and the proliferation rate of RA FLS was evaluated by EDU assays.Triad1 overexpression plasmid was used to enhance the expression of Triad1 in RA FLS.After transfection,the RA FLS were stimulated with ADR for 24 h,and the apoptosis of RA FLS was determined by EDU staining.In order to explore the pathway for Triad1-induced apoptosis of RA FLS,Western blot analysis was firstly employed to detect the possible function of p53 in Triad1-induced apoptosis of RA FLS.RA FLS were transfetcted with Triad1 overexpression plasmid to enhance the expression of Triad1,transfected RA FLS with p53-shRNA to attenuate p53 expreesion,and stimulated RA FLS with ADR to induce apoptosis.Several apoptosis related protein levels were detected under different transfection conditions.Finally,flow cytometry analysis was used to determine the possible role of p53 in Triad1-induced apoptosis of RA FLS.Results Triad1 was expressed in RA synovial tissues and had correlation with the apoptosis of RA FLS.Triad1 had negative effects on the proliferation of RA FLS.Triad1 induced the apoptosis of RA FLS likely via p53 pathway.Conclusions Triad1 might promote the apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes through p53 pathway.