JNK and PI3K differentially regulate MMP-2 and MT1-MMP mRNA and protein in response to actin cytoskeleton reorganization in endothelial cells
JNK and PI3K differentially regulate MMP-2 and MT1-MMP mRNA and protein in response to actin cytoskeleton reorganization in endothelial cells
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DOI：
10.1152/ajpcell.00300.2005
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发表时间：
2006-10-01
影响因子：
5.5
通讯作者：
Haas, Tara L.
中科院分区：
文献类型：
--
作者：
Ispanovic, Eric;Haas, Tara L.
基质金属蛋白酶-2（MMP-2）的产生和活化增加是骨骼肌血管生成中的关键事件，并且已知其响应于机械应力而发生。我们假设肌动蛋白细胞骨架的重组会增加内皮细胞MMP-2的产生和激活，并且这种增加需要内皮细胞中的MAPK依赖性信号通路。药理学肌动蛋白解聚剂细胞松弛素D增加MMP-2和膜型1-基质金属蛋白酶（MT 1-MMP）mRNA的表达，并在JNK抑制剂SP 600125的存在下显着降低。茴香霉素激活JNK足以诱导静止细胞中MMP-2和MT 1-MMP mRNA的表达。下调c-Jun（JNK的下游靶点），用小干扰（si）RNA抑制MMP-2的表达，以响应茴香霉素。磷酸肌醇3-激酶（PI 3 K）的抑制，但不是JNK，显着降低活性MMP-2的细胞松弛素D刺激后的MT 1-MMP蛋白的同时减少量。在VEGF刺激期间发生肌动蛋白的生理重组。VEGF诱导的MMP-2蛋白的产生和活化，以及MT 1-MMP蛋白的产生，依赖于PI 3 K活性。VEGF诱导的MMP-2 mRNA表达通过抑制JNK或用c-Jun siRNA治疗而降低。总之，我们的研究结果提供了新的见解信号级联在血管生成的早期阶段，通过重组肌动蛋白细胞骨架，并证明了一个关键的作用，JNK在调节MMP-2和MT 1-MMP mRNA的表达，而PI 3 K调节MMP-2和MT 1-MMP的蛋白水平。
Increased production and activation of matrix metalloproteinase-2 (MMP-2) are critical events in skeletal muscle angiogenesis and are known to occur in response to mechanical stresses. We hypothesized that reorganization of the actin cytoskeleton would increase endothelial cell production and activation of MMP-2 and that this increase would require a MAPK-dependent signaling pathway in endothelial cells. The pharmacological actin depolymerization agent cytochalasin D increased expression of MMP-2 and membrane type 1-matrix metalloproteinase (MT1-MMP) mRNA, and this was reduced significantly in the presence of the JNK inhibitor SP600125. Activation of JNK by anisomycin was sufficient to induce expression of both MMP-2 and MT1-MMP mRNA in quiescent cells. Downregulation of c-Jun, a downstream target of JNK, with small interference (si) RNA inhibited MMP-2 expression in response to anisomycin. Inhibition of phosphoinositide 3-kinase (PI3K), but not JNK, significantly decreased the amount of active MMP-2 following cytochalasin D stimulation with a concurrent decrease in MT1-MMP protein. Physiological reorganization of actin occurs during VEGF stimulation. VEGF-induced MMP-2 protein production and activation, as well as MT1-MMP protein production, depended on PI3K activity. VEGF-induced MMP-2 mRNA expression was reduced by inhibition of JNK or by treatment with c-Jun siRNA. In summary, our results provide novel insight into the signaling cascades initiated in the early stages of angiogenesis through the reorganization of the actin cytoskeleton and demonstrate a critical role for JNK in regulating MMP-2 and MT1-MMP mRNA expression, whereas PI3K regulates protein levels of both MMP-2 and MT1-MMP.