白细胞介素-1β通过丝裂原活化蛋白激酶/活化蛋白-1和核因子κB依赖性基质金属蛋白酶-9的表达促进A549细胞迁移。
IL-1 beta promotes A549 cell migration via MAPKs/AP-1- and NF-kappaB-dependent matrix metalloproteinase-9 expression.
作者信息
Lin Chih-Chung, Kuo Chang-Ting, Cheng Ching-Yi, Wu Cheng-Ying, Lee Chiang-Wen, Hsieh Hsi-Lung, Lee I-Ta, Yang Chuen-Mao
机构信息
Department of Anesthetics, Chang Gung University and Chang Gung Memorial Hospital, Kwei-San, Tao-Yuan, Taiwan, Republic of China.
出版信息
Cell Signal. 2009 Nov;21(11):1652-62. doi: 10.1016/j.cellsig.2009.07.002. Epub 2009 Jul 16.
Matrix metalloproteinases (MMPs), in particular MMP-9, is induced by cytokines including IL-1 beta and contributes to airway injury and remodeling. However, the mechanisms underlying IL-1 beta-induced MMP-9 expression and cell migration in human A549 cells remain unclear. Here, we report that the IL-1 beta-induced MMP-9 gene expression was mediated through the activation of p42/p44 MAPK, p38 MAPK, and JNK1/2 in A549 cells, determined by zymographic, RT-PCR, and Western blotting. The involvement of MAPKs in the IL-1beta-induced responses was further ensured by transfection with siRNA of MEK1, p42, p38, or JNK2. Moreover, the IL-1 beta-induced MMP-9 gene expression was also mediated through the translocation of NF-kappaB (p65) into the nucleus and the degradation of I kappaB alpha. In addition, the IL-1 beta-induced c-Jun phosphorylation was reduced by pretreatment with U0126 or SP600125. IL-1 beta stimulated the transcriptional activity of wild-type MMP-9 promoter in A549 cells, which was inhibited by U0126, SB203580, SP600125, and helenalin. In contrast, IL-1 beta had no effect on the cells transfected with a NF-kappaB-mutated MMP-9 promoter construct, suggesting that NF-kappaB is required for this response. Finally, the IL-1 beta-induced MMP-9 expression led to cell migration which was attenuated by pretreatment with U0126, SB203580, SP600125, helenalin, or MMP-2/9 inhibitor. These results suggested that in A549 cells, the activation of p42/p44 MAPK, p38 MAPK, JNK1/2, NF-kappaB, and AP-1 are essential for the IL-1 beta-induced MMP-9 gene expression and cell migration.
基质金属蛋白酶(MMPs),尤其是MMP-9,可由包括白细胞介素-1β(IL-1β)在内的细胞因子诱导产生,并参与气道损伤和重塑过程。然而,IL-1β诱导人A549细胞中MMP-9表达及细胞迁移的潜在机制仍不清楚。在此,我们报告,通过酶谱分析、逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法确定,IL-1β诱导的MMP-9基因表达是通过A549细胞中p42/p44丝裂原活化蛋白激酶(MAPK)、p38 MAPK和JNK1/2的激活介导的。通过用MEK1、p42、p38或JNK2的小干扰RNA(siRNA)转染进一步证实了MAPKs参与IL-1β诱导的反应。此外,IL-1β诱导的MMP-9基因表达还通过核因子-κB(NF-κB,p65)易位至细胞核以及IκBα降解介导。另外,用U0126或SP600125预处理可降低IL-1β诱导的c-Jun磷酸化。IL-1β刺激A549细胞中野生型MMP-9启动子的转录活性,这被U0126、SB203580、SP600125和海兔毒素抑制。相反,IL-1β对转染了NF-κB突变的MMP-9启动子构建体的细胞无影响,表明NF-κB是此反应所必需的。最后,IL-1β诱导的MMP-9表达导致细胞迁移,用U0126、SB203580、SP600125、海兔毒素或MMP-2/9抑制剂预处理可减弱这种迁移。这些结果表明,在A549细胞中,p42/p44 MAPK、p38 MAPK、JNK1/2、NF-κB和活化蛋白-1(AP-1)的激活对于IL-1β诱导的MMP-9基因表达和细胞迁移至关重要。