Direct recruitment of CRK and GRB2 to VEGFR-3 induces proliferation, migration, and survival of endothelial cells through the activation of ERK, AKT, and JNK pathways
Direct recruitment of CRK and GRB2 to VEGFR-3 induces proliferation, migration, and survival of endothelial cells through the activation of ERK, AKT, and JNK pathways
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DOI：
10.1182/blood-2005-04-1388
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发表时间：
2005-11-15
期刊：
影响因子：
20.3
通讯作者：
Oliviero, S
中科院分区：
文献类型：
--
作者：
Salameh, A;Galvagni, F;Oliviero, S
血管内皮生长因子受体-3(VEGFR-3)在胚胎初级毛细血管丛的重建中起关键作用，并在成人的血管生成和淋巴管生成中起重要作用。然而，VEGFR-3的信号转导途径仍有待阐明。在这里，我们通过系统地突变可能参与VEGFR-3信号转导的酪氨酸残基，研究了原代人脐静脉内皮细胞(HUVECs)中的VEGFR-3信号转导，并鉴定了对其功能至关重要的酪氨酸。研究表明，Y1068对受体的激酶活性是必需的。Y1063通过将CRKI/II募集到激活的受体上，诱导信号级联，通过丝裂原激活的蛋白激酶-4(MKK4)激活c-Jun氨基末端激酶1/2(JNK1/2)，从而发出受体介导的存活信号。特异性多肽抑制剂JNKI1或RNA干扰(RNAi)抑制JNK1/2的功能表明，JNK1/2的激活是VEGFR-3依赖的生存信号所必需的。Y1230/Y1231与Y1337共同促进内皮细胞的增殖、迁移和存活。磷酸化的Y1230/Y1231直接向受体募集生长因子受体结合蛋白(Grb2)，诱导AKT和细胞外信号相关蛋白1/2(ERK1/2)信号的激活。最后，我们观察到Y1063和Y11230/Y1231信号融合诱导c-jun的表达，RNAi实验证明c-jun是生长因子诱导的原代内皮细胞生存信号所必需的。
Vascular endothelial growth factor receptor-3 (VEGFR-3) plays a key role for the remodeling of the primary capillary plexus in the embryo and contributes to angiogenesis and lymphangiogenesis in the adult. However, VEGFR-3 signal transduction pathways remain to be elucidated. Here we investigated VEGFR-3 signaling in primary human umbilical vein endothelial cells (HUVECs) by the systematic mutation of the tyrosine residues potentially involved in VEGFR-3 signaling and identified the tyrosines critical for its function. Y1068 was shown to be essential for the kinase activity of the receptor. Y1063 Signals the receptor-mediated survival by recruiting CRKI/II to the activated receptor, inducing a signaling cascade that, via mitogen-activated protein kinase kinase-4 (MKK4), activates c-Jun N-terminal kinase1/2 (JNK1/2). Inhibition of JNK1/2 function either by specific peptide inhibitor JNKI1 or by RNA interference (RNAi) demonstrated that activation of JNK1/2 is required for a VEGFR-3-dependent prosurvival signaling. Y1230/Y1231 contributes, together with Y1337, to proliferation, migration, and survival of endothelial cells. Phospho-Y1230/Y1231 directly recruits growth factor receptor-bonus protein (GRB2) to the receptor, inducing the activation of both AKT and extracellular signal-related kinase 1/2 (ERK1/2) signaling. Finally, we observed that Y1063 and Y11230/ Y1231 signaling converge to induce c-JUN expression, and RNAi experiments demonstrated that c-JUN is required for growth factor-induced prosurvival signaling in primary endothelial cells.