IL-17A and IL-17F induce an invasive phenotype in human rheumatoid synoviocytes through the same signaling pathway and the CXCR4/SDF1 axis
IL-17A and IL-17F induce an invasive phenotype in human rheumatoid synoviocytes through the same signaling pathway and the CXCR4/SDF1 axis
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IL-17A 和 IL-17F 通过相同的信号通路和 CXCR4/SDF1 轴在人类风湿滑膜细胞中诱导侵袭表型
DOI：
10.1136/ard.2010.149104.14
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发表时间：
2011
影响因子：
27.4
通讯作者：
P. Miossec
中科院分区：
文献类型：
--
作者：
A. Hot;P. Miossec
目的探讨IL-17A与类风湿关节炎（RA）发病机制的关系。但IL-17F的作用仍有待澄清。本报告分析IL-17A与IL-17F对人RA滑膜细胞基因表达、信号传导和侵袭性的影响。方法采用微阵列技术（Affymetrix U133 +2）在mRNA水平上比较IL-17A和F对RA滑膜细胞的影响。采用Western blotting、qRT-PCR和DNA结合试验对其信号通路进行评价。利用transwell Matrigel侵袭室检测IL-17A和F单独或与TNF联合诱导滑膜细胞迁移和侵袭的能力。采用功能性DNA结合试验评价缺氧诱导因子1 （HIF1-α）活化的调控作用。结果在微阵列中，IL-17A和IL-17F单独具有非常相似但不完全相同的调节作用，IL-17F活性较低，在TNF存在时具有协同模式。这种协同效应部分与IL-17A和F增强TNF - RII的表达有关。至于它们的信号通路，几乎所有的IL-17A和F诱导基因都依赖于NF-κB的激活，而一小部分由p38调节。在缺氧诱导基因中，IL-17A和F单独或与TNF联合诱导的CXCR4 mRNA （IL-17A 289倍，IL-17F 34倍，IL-17A与TNF联合450倍）被IL-17A和F激活。IF染色证实了CXCR4在滑膜细胞表面的过表达。IL-17A和TNF通过CXCR4协同诱导滑膜细胞迁移和侵袭（6 vs 67个迁移细胞/HPF， p≤0.05）。阻断CXCR4可减少滑膜细胞迁移（- 10倍，p<0.05）。在常氧条件下，IL-17A或F与TNF联合可诱导HIF1-α活化（2倍，p=0.003）。结论IL-17A和F联合TNF诱导RA滑膜细胞表达模式相似但不完全相同，可能通过增加滑膜细胞侵袭性促进RA的进展。这种作用部分来自介导的CXCR4/SDF1途径。这些结果与IL-17A和F在RA中的靶向一致。
Objective IL-17A is implicated in rheumatoid arthritis (RA) pathogenesis; but the contribution of IL-17F remains to be clarified. This report analyses the effects IL-17A versus IL-17F on gene expression, signaling and invasiveness in human RA synoviocytes. Methods The comparison between IL-17A and F effects on RA synoviocytes was assessed at the mRNA level by microarrays (Affymetrix U133 +2). Western blotting, qRT-PCR, and DNA binding assay were used to evaluate their signaling pathways. The capacities of IL-17A and F alone or in combination with TNF to induce synoviocyte migration and invasion were tested using transwell Matrigel invasion chambers. A functional DNA binding assay was used to evaluate the regulation of Hypoxia Induced Factor 1 (HIF1-α) activation. Results In microarrays, IL-17A and IL-17F alone had very similar but not identical regulatory effects, IL-17F being less active, with a synergistic pattern in the presence of TNF. This synergistic effect was linked in part to the enhancing expression of TNF RII by IL-17A and F. Regarding their signaling pathway, virtually all IL17A and F inducible genes were dependent on NF-κB activation, whereas a minor number was modulated by p38. Hypoxia-induced pathway was activated by IL-17A and F. Among the hypoxia-induced genes, IL-17A and F alone or combined with TNF induced CXCR4 mRNA (289 fold for IL17A, 34 fold for IL-17F, 450 fold for the combination of IL-17A with TNF). Over expression of CXCR4 at the surface of synoviocytes was confirmed by IF staining. IL-17A and TNF induced in synergy synoviocyte migration and invasion through CXCR4 (6 vs 67 migrated cells/HPF, p≤0.05). Blockade of CXCR4 decreased synoviocyte migration (− 10 fold, p<0.05). The combination of IL-17A or F with TNF induced activation of HIF1-α under normoxic conditions (2 fold, p=0.003). Conclusion When combined with TNF, IL-17A and F induced very similar but not identical expression pattern in RA synoviocytes, may contribute to the progression of RA through increased synoviocyte aggressiveness. Part of this effect results from a mediated CXCR4/SDF1 pathway. These results are in line with IL-17A and F targeting in RA.