Early growth response factor-1 mediates prostaglandin E2-dependent transcriptional suppression of cytokine-induced tumor necrosis factor-α gene expression in human macrophages and rheumatoid arthritis-affected synovial fibroblasts
Early growth response factor-1 mediates prostaglandin E2-dependent transcriptional suppression of cytokine-induced tumor necrosis factor-α gene expression in human macrophages and rheumatoid arthritis-affected synovial fibroblasts
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DOI：
10.1074/jbc.m414067200
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发表时间：
2005-03-11
影响因子：
4.8
通讯作者：
Di Battista, JA
中科院分区：
文献类型：
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作者：
Faour, WH;Alaaeddine, N;Di Battista, JA
肿瘤坏死因子- α (tnf - α)是一种多效性促炎细胞因子，可调节广泛的炎症和免疫过程。我们研究了前列腺素E-2 （PGE(2)）抑制人巨噬细胞和类风湿关节炎影响的滑膜成纤维细胞中t细胞来源的白介素-17 （IL-17）诱导的tnf - α mRNA表达和蛋白质合成的分子机制，从而研究了PGE(2)的潜在免疫调节特性。初步研究证实，通过野生型（WT）和缺失突变型egr-1启动子构建、Northern和Western blotting以及标准和超移电泳迁移迁移分析，PGE2通过限制SAPK2/p38 mapk依赖性激活转录因子-2 （ATF-2）二聚体反激活egr-1启动子，PGE2诱导egr-1 mRNA表达和蛋白质合成。使用稳定转染WT和Egr-1显性阴性突变构建体的人白血病单核THP-1细胞，无论是否共转染WT pTNF-615SVOCAT构建体，我们观察到PGE2对il -17刺激的tnf - α mRNA表达和启动子活性的抑制依赖于Egr-1的表达，因为Egr-1突变体单独或联合显着消除了PGE2的任何抑制作用。标准和超移电泳迁移率转移分析、信号“诱饵”过表达研究以及使用WT和突变启动子构建的pTNF-615SVOCAT启动子分析显示，il -17上调的启动子活性在很大程度上依赖于ATF-2/c-Jun的转激活。PGE(2)对il -17诱导的atf -2/c-Jun反活化和DNA结合的抑制依赖于egr -1介导的对诱导c-Jun表达的抑制。我们认为egr-1是一种早期PGE靶基因，可能是介导免疫和炎症反应相关基因的类二十烷控制的关键调节因子。
Tumor necrosis factor-alpha (TNF-alpha) is a pleiotropic pro-inflammatory cytokine that modulates a broad range of inflammatory and immunological processes. We have investigated the potential immunomodulatory properties of prostaglandin E-2 (PGE(2)) by examining the molecular mechanism by which the eicosanoid suppresses T-cell-derived interleukin-17 (IL-17)-induced TNF-alpha mRNA expression and protein synthesis in human macrophages and rheumatoid arthritis-affected synovial fibroblasts. Initial studies confirmed that PGE2 induces egr-1 mRNA expression and protein synthesis by restricted SAPK2/p38 MAPK-dependent activating transcription factor-2 (ATF-2) dimer transactivation of the egr-1 promoter as judged by studies using wild-type (WT) and deletion mutant egr-1 promoter constructs, Northern and Western blotting, and standard and supershift electrophoretic mobility shift analyses. Using human leukemic monocytic THP-1 cells stably transfected with WT and dominant-negative mutant expression constructs of Egr-1, cotransfected or not with a WT pTNF-615SVOCAT construct, we observed that PGE2 inhibition of IL-17-stimulated TNF-alpha mRNA expression and promoter activity was dependent on Egr-1 expression, as mutants of Egr-1, alone or in combination, markedly abrogated any inhibitory effect of PGE2. Standard and supershift electrophoretic mobility shift analysis, signaling "decoy" overexpression studies, and pTNF-615SVOCAT promoter assays using WT and mutant promoter constructs revealed that IL-17-up-regulated promoter activity was largely dependent on ATF-2/c-Jun transactivation. PGE(2) suppression of IL-17-inducedATF-2/c-Jun transactivation and DNA binding was dependent on Egr-1-mediated inhibition of induced c-Jun expression. We suggest that egr-1 is an immediate-early PGE(2) target gene that may be a key regulatory factor in mediating eicosanoid control of genes involved in the immune and inflammatory responses.