Decoy receptor 3 expressed in rheumatoid synovial fibroblasts protects the cells against fas-induced apoptosis
Decoy receptor 3 expressed in rheumatoid synovial fibroblasts protects the cells against fas-induced apoptosis
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DOI：
10.1002/art.22494
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发表时间：
2007-04-01
影响因子：
--
通讯作者：
Doita, Minoru
中科院分区：
文献类型：
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作者：
Hayashi, Shinya;Miura, Yasushi;Doita, Minoru
目标。诱饵受体3 （DcR3）是新发现的肿瘤坏死因子受体（TNFR）超家族成员，是一种可与TNF家族成员结合的可溶性受体，包括FasL、LIGHT和TNF样分子1A。DcR3主要在肿瘤细胞中表达，它竞争性地抑制TNF与tnfr的结合。本研究旨在研究DcR3在类风湿关节炎（RA）和骨关节炎（OA）患者成纤维细胞样滑膜细胞（FLS）中的表达，并分析DcR3对fas诱导的类风湿关节炎（RA）成纤维细胞样滑膜细胞凋亡的影响。采用逆转录聚合酶链反应（RT-PCR）和Western blotting检测FLS中DcR3的表达。用DcR3- fc嵌合体蛋白孵育FLS或用脂质体法转染DcR3小干扰RNA (siRNA)，诱导FLS凋亡。采用TUNEL染色和caspase 8、聚adp核糖聚合酶Western blotting检测Fas诱导的FLS凋亡。最后，在fas诱导细胞凋亡前，将FLS与TNF - α孵育，采用定量RT-PCR分析DcR3的表达，并检测细胞凋亡。DcR3在RA FLS和OA FLS中均有表达。DcR3-Fc蛋白抑制fas诱导的FLS细胞凋亡。siRNA下调FLS中的DcR3可增加fas诱导的细胞凋亡。TNF α在RA FLS中升高DcR3表达，抑制fas诱导的细胞凋亡，而在OA FLS中无此作用。DcR3在RA FLS中的表达被TNF α增加，并保护细胞免受fas诱导的凋亡。这些发现表明DcR3可能是RA的一个可能的治疗靶点。
Objective. Decoy receptor 3 (DcR3), a newly identified member of the tumor necrosis factor receptor (TNFR) superfamily, is a soluble receptor that binds to members of the TNF family, including FasL, LIGHT, and TNF-like molecule 1A. DcR3 is mostly expressed in tumor cells, and it competitively inhibits binding of TNF to TNFRs. The present study was undertaken to investigate DcR3 expression in fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA) and osteoarthritis (OA), and to analyze the effects of DcR3 on Fas-induced apoptosis in RA FLS.Methods. Expression of DcR3 in FLS was measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. FLS were incubated with DcR3-Fc chimera protein or transfected with DcR3 small interfering RNA (siRNA) using the lipofection method, before induction of apoptosis. Apoptosis induced by Fas in FLS was detected with TUNEL staining and Western blotting of caspase 8 and poly(ADP-ribose) polymerase. Finally, FLS were incubated with TNF alpha prior to Fas-induced apoptosis, expression of DcR3 was analyzed by quantitative RT-PCR, and apoptosis was measured.Results. DcR3 was expressed in both RA FLS and OA FLS. DcR3-Fc protein inhibited Fas-induced apoptosis in FLS. Down-regulation of DcR3 in FLS by siRNA increased Fas-induced apoptosis. TNF alpha increased DcR3 expression and inhibited Fas-induced apoptosis in RA FLS, but not in OA FLS.Conclusion. DcR3 expressed in RA FLS is increased by TNF alpha and protects the cells against Fas-induced apoptosis. These findings indicate that DcR3 may be a possible therapeutic target in RA.