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Activation of vascular endothelial growth factor receptor-3 and its downstream signaling promote cell survival under oxidative stress.

Identifieur interne : 001974 ( Ncbi/Curation ); précédent : 001973; suivant : 001975

Activation of vascular endothelial growth factor receptor-3 and its downstream signaling promote cell survival under oxidative stress.

Auteurs : Jian Feng Wang [États-Unis] ; Xuefeng Zhang ; Jerome E. Groopman

Source :

RBID : pubmed:15102829

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English descriptors

Abstract

Reactive oxygen species (ROS) mediate cell damage and have been implicated in the pathogenesis of diseases that involve endothelial injury. Cells possess antioxidant systems, including intracellular antioxidants and ROS scavenging enzymes, that control the redox state and prevent cell damage. In addition to intracellular antioxidants, certain growth factor receptors can be activated under oxidative stress and trigger downstream cell survival signaling cascades. Vascular endothelial growth factor receptor-3 (VEGFR-3) is a primary modulator of lymphatic endothelial proliferation and survival. Here, we provide evidence that activation of VEGFR-3 signaling in response to hydrogen peroxide (H(2)O(2)) promotes endothelial cell survival. Treatment with H(2)O(2) induced the tyrosine phosphorylation of VEGFR-3 and its association with the signaling adaptor proteins Shc, growth factor receptor binding protein 2, Sos, p85, SHP-2, and phospholipase C-gamma. Of note, a hereditary lymphoedema-linked mutant of VEGFR-3 was not phosphorylated by H(2)O(2) treatment. Isoforms of protein kinase C (PKC), alpha and delta, were also tyrosine-phosphorylated after H(2)O(2) stimulation. However, only the delta isoform of PKC was required for H(2)O(2)-induced phosphorylation of VEGFR-3. The tyrosine phosphorylation of VEGFR-3 or isoforms of PKC was completely inhibited by treatment with 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine, a specific inhibitor for Src family kinases, indicating that Src family kinases are upstream of PKC and VEGFR-3. Furthermore, expression of the wild-type but not the lymphoedema-linked mutant form of VEGFR-3 in porcine artery endothelial cells significantly enhanced the activation of Akt after H(2)O(2) stimulation. Consistent with these biochemical changes, we observed that expression and activation of the wild-type but not the mutant form of VEGFR-3 inhibited H(2)O(2)-induced apoptosis. These studies suggest that VEGFR-3 protects against oxidative damage in endothelial cells, and that patients with hereditary lymphoedema may be susceptible to ROS-induced cell damage.

DOI: 10.1074/jbc.M314015200
PubMed: 15102829

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<term>Cell Line</term>
<term>Cell Survival (physiology)</term>
<term>Endothelium, Vascular (cytology)</term>
<term>Enzyme Activation (drug effects)</term>
<term>Enzyme Inhibitors (pharmacology)</term>
<term>Humans</term>
<term>Hydrogen Peroxide (pharmacology)</term>
<term>Lymphedema (genetics)</term>
<term>Lymphedema (metabolism)</term>
<term>Mutation, Missense</term>
<term>Oncogene Protein v-akt</term>
<term>Oxidative Stress (drug effects)</term>
<term>Oxidative Stress (physiology)</term>
<term>Phosphoric Monoester Hydrolases (antagonists & inhibitors)</term>
<term>Phosphorylation</term>
<term>Protein Kinase C (antagonists & inhibitors)</term>
<term>Protein Kinase C (genetics)</term>
<term>Protein Kinase C (metabolism)</term>
<term>Pyrimidines (pharmacology)</term>
<term>Recombinant Proteins (genetics)</term>
<term>Recombinant Proteins (metabolism)</term>
<term>Retroviridae Proteins, Oncogenic (metabolism)</term>
<term>Signal Transduction</term>
<term>Tyrosine (metabolism)</term>
<term>Vanadates (pharmacology)</term>
<term>Vascular Endothelial Growth Factor Receptor-3 (genetics)</term>
<term>Vascular Endothelial Growth Factor Receptor-3 (metabolism)</term>
<term>Vascular Endothelial Growth Factor Receptor-3 (physiology)</term>
<term>src Homology Domains</term>
<term>src-Family Kinases (antagonists & inhibitors)</term>
<term>src-Family Kinases (genetics)</term>
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<term>Activation enzymatique ()</term>
<term>Antienzymes (pharmacologie)</term>
<term>Domaine d'homologie SRC</term>
<term>Endothélium vasculaire (cytologie)</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Lymphoedème (génétique)</term>
<term>Lymphoedème (métabolisme)</term>
<term>Mutation faux-sens</term>
<term>Peroxyde d'hydrogène (pharmacologie)</term>
<term>Phosphoric monoester hydrolases (antagonistes et inhibiteurs)</term>
<term>Phosphorylation</term>
<term>Protéine kinase C (antagonistes et inhibiteurs)</term>
<term>Protéine kinase C (génétique)</term>
<term>Protéine kinase C (métabolisme)</term>
<term>Protéine oncogène v-akt</term>
<term>Protéines oncogènes des retroviridae (métabolisme)</term>
<term>Protéines recombinantes (génétique)</term>
<term>Protéines recombinantes (métabolisme)</term>
<term>Pyrimidines (pharmacologie)</term>
<term>Récepteur-3 au facteur croissance endothéliale vasculaire (génétique)</term>
<term>Récepteur-3 au facteur croissance endothéliale vasculaire (métabolisme)</term>
<term>Récepteur-3 au facteur croissance endothéliale vasculaire (physiologie)</term>
<term>Stress oxydatif ()</term>
<term>Stress oxydatif (physiologie)</term>
<term>Survie cellulaire (physiologie)</term>
<term>Transduction du signal</term>
<term>Tyrosine (métabolisme)</term>
<term>Vanadates (pharmacologie)</term>
<term>src-Family kinases (antagonistes et inhibiteurs)</term>
<term>src-Family kinases (génétique)</term>
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<term>Phosphoric Monoester Hydrolases</term>
<term>Protein Kinase C</term>
<term>src-Family Kinases</term>
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<term>Protein Kinase C</term>
<term>Recombinant Proteins</term>
<term>Vascular Endothelial Growth Factor Receptor-3</term>
<term>src-Family Kinases</term>
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<term>Protein Kinase C</term>
<term>Recombinant Proteins</term>
<term>Retroviridae Proteins, Oncogenic</term>
<term>Tyrosine</term>
<term>Vascular Endothelial Growth Factor Receptor-3</term>
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<term>Enzyme Inhibitors</term>
<term>Hydrogen Peroxide</term>
<term>Pyrimidines</term>
<term>Vanadates</term>
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<term>Phosphoric monoester hydrolases</term>
<term>Protéine kinase C</term>
<term>src-Family kinases</term>
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<term>Endothélium vasculaire</term>
</keywords>
<keywords scheme="MESH" qualifier="cytology" xml:lang="en">
<term>Endothelium, Vascular</term>
</keywords>
<keywords scheme="MESH" qualifier="drug effects" xml:lang="en">
<term>Enzyme Activation</term>
<term>Oxidative Stress</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en">
<term>Lymphedema</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr">
<term>Lymphoedème</term>
<term>Protéine kinase C</term>
<term>Protéines recombinantes</term>
<term>Récepteur-3 au facteur croissance endothéliale vasculaire</term>
<term>src-Family kinases</term>
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<keywords scheme="MESH" qualifier="metabolism" xml:lang="en">
<term>Lymphedema</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr">
<term>Lymphoedème</term>
<term>Protéine kinase C</term>
<term>Protéines oncogènes des retroviridae</term>
<term>Protéines recombinantes</term>
<term>Récepteur-3 au facteur croissance endothéliale vasculaire</term>
<term>Tyrosine</term>
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<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr">
<term>Antienzymes</term>
<term>Peroxyde d'hydrogène</term>
<term>Pyrimidines</term>
<term>Vanadates</term>
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<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr">
<term>Récepteur-3 au facteur croissance endothéliale vasculaire</term>
<term>Stress oxydatif</term>
<term>Survie cellulaire</term>
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<term>Cell Survival</term>
<term>Oxidative Stress</term>
<term>Vascular Endothelial Growth Factor Receptor-3</term>
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<keywords scheme="MESH" xml:lang="en">
<term>Cell Line</term>
<term>Humans</term>
<term>Mutation, Missense</term>
<term>Oncogene Protein v-akt</term>
<term>Phosphorylation</term>
<term>Signal Transduction</term>
<term>src Homology Domains</term>
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<term>Activation enzymatique</term>
<term>Domaine d'homologie SRC</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Mutation faux-sens</term>
<term>Phosphorylation</term>
<term>Protéine oncogène v-akt</term>
<term>Stress oxydatif</term>
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<div type="abstract" xml:lang="en">Reactive oxygen species (ROS) mediate cell damage and have been implicated in the pathogenesis of diseases that involve endothelial injury. Cells possess antioxidant systems, including intracellular antioxidants and ROS scavenging enzymes, that control the redox state and prevent cell damage. In addition to intracellular antioxidants, certain growth factor receptors can be activated under oxidative stress and trigger downstream cell survival signaling cascades. Vascular endothelial growth factor receptor-3 (VEGFR-3) is a primary modulator of lymphatic endothelial proliferation and survival. Here, we provide evidence that activation of VEGFR-3 signaling in response to hydrogen peroxide (H(2)O(2)) promotes endothelial cell survival. Treatment with H(2)O(2) induced the tyrosine phosphorylation of VEGFR-3 and its association with the signaling adaptor proteins Shc, growth factor receptor binding protein 2, Sos, p85, SHP-2, and phospholipase C-gamma. Of note, a hereditary lymphoedema-linked mutant of VEGFR-3 was not phosphorylated by H(2)O(2) treatment. Isoforms of protein kinase C (PKC), alpha and delta, were also tyrosine-phosphorylated after H(2)O(2) stimulation. However, only the delta isoform of PKC was required for H(2)O(2)-induced phosphorylation of VEGFR-3. The tyrosine phosphorylation of VEGFR-3 or isoforms of PKC was completely inhibited by treatment with 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine, a specific inhibitor for Src family kinases, indicating that Src family kinases are upstream of PKC and VEGFR-3. Furthermore, expression of the wild-type but not the lymphoedema-linked mutant form of VEGFR-3 in porcine artery endothelial cells significantly enhanced the activation of Akt after H(2)O(2) stimulation. Consistent with these biochemical changes, we observed that expression and activation of the wild-type but not the mutant form of VEGFR-3 inhibited H(2)O(2)-induced apoptosis. These studies suggest that VEGFR-3 protects against oxidative damage in endothelial cells, and that patients with hereditary lymphoedema may be susceptible to ROS-induced cell damage.</div>
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