The angiogenic regulator CD13/APN is a transcriptional target of Ras signaling pathways in endothelial morphogenesis
The angiogenic regulator CD13/APN is a transcriptional target of Ras signaling pathways in endothelial morphogenesis
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DOI：
10.1182/blood-2002-05-1422
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发表时间：
2003-03-01
期刊：
影响因子：
20.3
通讯作者：
Shapiro, LH
中科院分区：
文献类型：
--
作者：
Bhagwat, SV;Petrovic, N;Shapiro, LH
血管生成，即新血管的形成，是肿瘤生长和转移的关键步骤，也是关节炎和增殖性视网膜病变病理炎症反应的组成部分。CD13/氨基肽酶N(CD13/APN)金属蛋白酶是血管生成的重要调节因子，其在激活的血管上的表达是由血管生成信号诱导的。在这里，我们证明了细胞因子诱导内皮细胞CD13/APN受不同的RAS效应通路调节，涉及RAS/丝裂原活化蛋白激酶(MAPK)或PI-3K。激活的RAS、大鼠和丝裂原诱导的细胞外激酶(MEK)转导的信号刺激CD13/APN近端启动子的转录。这些通路和细胞外信号调节的丝氨酸/苏氨酸激酶(ERK-2)和PI-3K的表达被显性负性蛋白或化学抑制剂抑制，从而阻止了碱性成纤维细胞生长因子(BFGF)对CD13/APN转录的诱导。我们发现，RAS诱导的信号转导是生长因子诱导的血管生成所必需的，因为抑制RAS信号的下游介质(MEK或PI-3K)在体外抑制内皮细胞的迁移、侵袭和形态形成。CD13/APN是这些通路的共同下游靶点，它的重新引入推翻了这些抑制物的抑制作用，并恢复了内皮细胞在迁移/侵袭和毛细血管形态发生分析中的功能。同样，抑制。在体内，MEK抑制了细胞的侵袭和内皮内毛细血管的形成，外源性CD13/APN蛋白可以有效地挽救这一过程。这些研究提供了强有力的证据表明，CD13/APN是RAS信号在血管生成中的重要靶点，并且是血管生成过程中的限制因素。
Angiogenesis, the formation of new blood vessels, is a critical step for tumor growth and metastasis and an integral component of the pathologic inflammatory response in arthritis and the proliferative retinopathies. The CD13/aminopeptidase N (CD13/APN) metalloprotease is an important regulator of angiogenesis where its expression on activated blood vessels is induced by angiogenic signals. Here, we show that cytokine induction of CD13/APN in endothelial cells is regulated by distinct Ras effector pathways involving Ras/mitogen-activated protein kinase (MAPK) or PI-3K. Signals transduced by activated Ras, Rat, and mitogen-induced extracellular kinase (MEK) stimulate transcription from the CD13/APN proximal promoter. Inhibition of these pathways and extracellular signal-regulated serine/threonine kinase (ERK-2) and PI-3K by expression of dominant-negative proteins or chemical inhibitors prevented induction of CD13/APN transcription in response to basic fibroblast growth factor (bFGF). We show that Ras-induced signal transduction is required for growth factor-induced angiogenesis, because inhibition of downstream mediators of Ras signaling (MEK or PI-3K) abrogated endothelial cell migration, invasion, and morphogenesis in vitro. Reintroduction of CD13/APN, a shared downstream target of these pathways, overrode the suppressive effect of these inhibitors and restored the function of endothelial cells in migration/invasion and capillary morphogenesis assays. Similarly, inhibition. of MEK abrogated cell invasion and the formation of endothelial-lined capillaries in vivo, which was effectively rescued by addition of exogenous CD13/APN protein. These studies provide strong evidence that CD13/APN is an important target of Ras signaling in angiogenesis and is a limiting factor in angiogenic progression.