ADAMTS-9 is synergistically induced by interleukin-1beta and tumor necrosis factor alpha in OUMS-27 chondrosarcoma cells and in human chondrocytes.
ADAMTS-9 is synergistically induced by interleukin-1beta and tumor necrosis factor alpha in OUMS-27 chondrosarcoma cells and in human chondrocytes.
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发表时间：
2005
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通讯作者：
K. Demircan;S. Hirohata;K. Nishida;O. Hatipoglu;T. Oohashi;T. Yonezawa;S. Apte;Y. Ninomiya
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作者：
K. Demircan;S. Hirohata;K. Nishida;O. Hatipoglu;T. Oohashi;T. Yonezawa;S. Apte;Y. Ninomiya
目的 比较软骨细胞样 OUMS-27 细胞和人软骨细胞中白细胞介素 1β (IL-1β) 和肿瘤坏死因子 α (TNFα) 对聚集蛋白聚糖酶 (ADAMTS-1、ADAMTS-4、ADAMTS-5、ADAMTS-8、ADAMTS-9 和 ADAMTS-15) 的诱导作用，并确定最敏感软骨细胞的诱导机制。聚集蛋白聚糖酶基因。方法用不同浓度的IL-1β和/或TNFα刺激OUMS-27细胞不同的时间。从骨关节炎关节获得的人软骨细胞和人皮肤成纤维细胞也用 IL-1β 和/或 TNFα 刺激。提取总RNA，逆转录，并通过定量实时聚合酶链反应和Northern印迹进行分析。通过蛋白质印迹法检查 ADAMTS-9 蛋白，并研究了 MAPK 信号通路在 IL-1β 刺激的 OUMS-27 细胞中诱导 ADAMTS9 的作用。结果 IL-1β 增加 ADAMTS4、ADAMTS5 和 ADAMTS9 的信使 RNA (mRNA) 水平，但不增加 ADAMTS1 和 ADAMTS8。 ADAMTS9 mRNA 的倍数增加大于其他聚集蛋白聚糖酶基因的 mRNA。 IL-1β 刺激引起的 ADAMTS9 mRNA 增加在软骨细胞中比在成纤维细胞中更大。 IL-1β和TNFα的组合具有协同作用，导致ADAMTS9 mRNA水平显着升高。 ADAMTS-9 蛋白也在 IL-1β 刺激的 OUMS-27 细胞中被诱导。 MAPK 抑制剂 SB203580 和 PD98059 降低了 OUMS-27 细胞中 ADAMTS9 的上调。结论 ADAMTS9 是一种 IL-1β 和 TNFα 诱导基因，与其他聚集蛋白聚糖酶基因相比，它似乎对这些促炎细胞因子更敏感。此外，这些细胞因子对 ADAMTS9 具有协同作用。连同 ADAMTS-9 已知的蛋白水解降解聚集蛋白聚糖的能力及其切割其他软骨分子的潜力，数据表明 ADAMTS-9 可能在关节炎中具有病理作用。
OBJECTIVE To compare induction of the aggrecanases (ADAMTS-1, ADAMTS-4, ADAMTS-5, ADAMTS-8, ADAMTS-9, and ADAMTS-15) by interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) in chondrocyte-like OUMS-27 cells and human chondrocytes, and to determine the mechanism of induction of the most responsive aggrecanase gene. METHODS OUMS-27 cells were stimulated for different periods of time and with various concentrations of IL-1beta and/or TNFalpha. Human chondrocytes obtained from osteoarthritic joints and human skin fibroblasts were also stimulated with IL-1beta and/or TNFalpha. Total RNA was extracted, reverse transcribed, and analyzed by quantitative real-time polymerase chain reaction and Northern blotting. ADAMTS-9 protein was examined by Western blotting, and the role of the MAPK signaling pathway for ADAMTS9 induction in IL-1beta-stimulated OUMS-27 cells was investigated. RESULTS IL-1beta increased messenger RNA (mRNA) levels of ADAMTS4, ADAMTS5, and ADAMTS9 but not ADAMTS1 and ADAMTS8. The fold increase for ADAMTS9 mRNA was greater than that for mRNA of the other aggrecanase genes. The increase of ADAMTS9 mRNA by IL-1beta stimulation was greater in chondrocytes than in fibroblasts. The combination of IL-1beta and TNFalpha had a synergistic effect, resulting in a considerable elevation in the level of ADAMTS9 mRNA. ADAMTS-9 protein was also induced in IL-1beta-stimulated OUMS-27 cells. The MAPK inhibitors SB203580 and PD98059 decreased ADAMTS9 up-regulation in OUMS-27 cells. CONCLUSION ADAMTS9 is an IL-1beta- and TNFalpha-inducible gene that appears to be more responsive to these proinflammatory cytokines than are other aggrecanase genes. Furthermore, these cytokines had a synergistic effect on ADAMTS9. Together with the known ability of ADAMTS-9 to proteolytically degrade aggrecan and its potential to cleave other cartilage molecules, the data suggest that ADAMTS-9 may have a pathologic role in arthritis.