IL-1β induces expression of matrix metalloproteinase-9 and cell migration via a c-Src-dependent, growth factor receptor transactivation in A549 cells
IL-1β induces expression of matrix metalloproteinase-9 and cell migration via a c-Src-dependent, growth factor receptor transactivation in A549 cells
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DOI：
10.1111/j.1476-5381.2010.00858.x
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发表时间：
2010-08-01
影响因子：
7.3
通讯作者：
Yang, Chuen-Mao
中科院分区：
文献类型：
--
作者：
Cheng, Ching-Yi;Kuo, Chang-Ting;Yang, Chuen-Mao
背景与目的白介素(IL)-1 β诱导的基质金属蛋白酶（MMP-9）的表达受丝裂原活化蛋白激酶（MAPKs）和NF-kappa b的调控，IL-1 β还刺激生长因子受体和磷脂酰肌醇3-激酶(PI3K)/Akt的反激活。，导致炎症蛋白的表达。在这里，我们研究了这些反激活机制是否参与IL-1 β诱导的A549细胞中MMP-9的表达。实验方法：在IL-1 β刺激前，用/不加药物抑制剂和中和抗体处理acha549细胞，或转染显性阴性突变体和特定蛋白激酶的siRNA。体外划痕法测定细胞迁移量。Western blot和明胶酶谱法分析MMP-9的表达和酶活性。采用RT-PCR、染色质免疫沉淀法和启动子法分析MMP-9的转录活性。关键结果通过Src （PP1）、血小板衍生生长因子（PDGF）受体和上皮生长因子（EGF）受体抑制剂或转染Src和Akt的siRNA抑制MMP-9的表达可阻止IL-1 β诱导的A549细胞迁移。这些酪氨酸激酶通过Src/PDGFR或Src/EGFR复合物的形成磷酸化Src、PDGF或EGF受体（EGFR）参与，这些复合物被PP1减弱。IL-1 β通过EGFR反激活诱导的MMP-9表达被MMPs和肝素结合egf样因子（HB-EGF）抑制剂或中和HB-EGF抗体降低。IL-1 β刺激Akt和NF-kappa B的激活和易位（p65）；LY294002减少了活化的NF-kappa B （p65）在MMP-9启动子区域的募集。结论和意义：在A549细胞中，sil -1 β诱导的MMP-9表达和细胞迁移是通过与NF-kappa B通路相关的EGFR/PDGFR/PI3K/Akt的c- src依赖性转激活介导的。
BACKGROUND AND PURPOSEInterleukin (IL)-1 beta-induced matrix metalloproteinase (MMP-9) expression is regulated by mitogen activated protein kinases (MAPKs) and NF-kappa B. IL-1 beta also stimulates transactivation of growth factor receptors and phosphatidylinositol 3-kinase (PI3K)/Akt., leading to the expression of inflammatory proteins. Here, we investigated whether these transactivation mechanisms participated in IL-1 beta-induced MMP-9 expression in A549 cells.EXPERIMENTAL APPROACHA549 cells were treated with/without pharmacological inhibitors and neutralizing antibody or transfected with dominant negative mutants and siRNA of particular protein kinases before stimulation with IL-1 beta. Cell migration was measured by in vitro scratch assay. Expression and enzymatic activity of MMP-9 were analysed by Western blot and gelatin zymography. Transcriptional activity of MMP-9 was analysed by RT-PCR, chromatin immunoprecipitation and promoter assays.KEY RESULTSInhibition of MMP-9 expression by inhibitors of Src (PP1), platelet-derived growth factor (PDGF) receptor and epithelial growth factor (EGF) receptor or transfection with siRNA for Src and Akt prevented IL-1 beta-induced migration of A549 cells. These tyrosine kinases were involved through phosphorylation of Src, PDGF, or EGF receptors (EGFRs) via the formation of Src/PDGFR or Src/EGFR complexes, attenuated by PP1. IL-1 beta-induced MMP-9 expression through EGFR transactivation was diminished by inhibitors of MMPs and heparin-binding EGF-like factor (HB-EGF), or a neutralizing HB-EGF antibody. IL-1 beta-stimulated activation and translocation of Akt and NF-kappa B (p65); the recruitment of activated NF-kappa B (p65) to the MMP-9 promoter region was attenuated by LY294002.CONCLUSIONS AND IMPLICATIONSIL-1 beta-induced MMP-9 expression and cell migration was mediated through c-Src-dependent transactivation of EGFR/PDGFR/PI3K/Akt linking to the NF-kappa B pathway in A549 cells.