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Antipermeability function of PEDF involves blockade of the MAP kinase/GSK/beta-catenin signaling pathway and uPAR expression.

ABSTRACT: Pigment epithelium-derived factor (PEDF) is a potent inhibitor of vascular endothelial growth factor (VEGF)-induced endothelial permeability. The goal of this study was to understand the mechanism by which PEDF blocks VEGF-induced increases in vascular permeability.The paracellular permeability of bovine retinal endothelial (BRE) cells was measured by assaying transendothelial cell electrical resistance and tracer flux. Western blot analysis was used to show phosphorylation of VEGFR2, MAP kinases, and glycogen synthase kinase 3 (GSK3)-beta. Confocal imaging and Western blot analysis were used to determine subcellular distribution of beta-catenin. Real-time RT-PCR and Western blot analysis were used to quantify urokinase plasminogen activator receptor (uPAR) expression.PEDF blocked VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAP kinase, the p38 substrate MAP kinase-activated protein kinase-2 (MAPKAPK-2), and GSK3-beta, but it had no effect on the phosphorylation of VEGFR2. In addition, the VEGF-induced transcriptional activation of beta-catenin and uPAR expression were blocked by PEDF and by inhibitors of p38 and MEK. Finally, the VEGF-induced increase in permeability was blocked by both PEDF and the same kinase inhibitors.The data suggest that p38 MAP kinase and ERK act upstream of GSK/beta-catenin in VEGF-induced activation of the uPA/uPAR system and that PEDF-mediated inhibition of the VEGF-induced increase in vascular permeability involves blockade of this pathway. These findings are important for developing precise and potent therapies for treatment of diseases characterized by vascular barrier dysfunction.

SUBMITTER: Yang J

PROVIDER: S-EPMC2891479 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature

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Antipermeability function of PEDF involves blockade of the MAP kinase/GSK/beta-catenin signaling pathway and uPAR expression.

Yang Jinling J Duh Elia J EJ Caldwell Ruth B RB Behzadian M Ali MA

Investigative ophthalmology & visual science 20100120 6

<h4>Purpose</h4>Pigment epithelium-derived factor (PEDF) is a potent inhibitor of vascular endothelial growth factor (VEGF)-induced endothelial permeability. The goal of this study was to understand the mechanism by which PEDF blocks VEGF-induced increases in vascular permeability.<h4>Methods</h4>The paracellular permeability of bovine retinal endothelial (BRE) cells was measured by assaying transendothelial cell electrical resistance and tracer flux. Western blot analysis was used to show phosp ...[more]

PMID: 20089873

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Project description:Fifty percent of cutaneous melanomas are driven by activated BRAFV600E, but tumors treated with RAF inhibitors, even when they respond dramatically, rapidly adapt and develop resistance. Thus, there is a pressing need to identify the major mechanisms of intrinsic and adaptive resistance and develop drug combinations that target these resistance mechanisms. In a combinatorial drug screen on a panel of 12 treatment-naïve BRAFV600E mutant melanoma cell lines of varying levels of resistance to MAPK pathway inhibition we identified the combination PLX4720, a targeted inhibitor of mutated BRaf, and lapatinib, an inhibitor of the ERBB family of receptor tyrosine kinases, as synergistically cytotoxic in the subset of cell lines that displayed the most resistance to PLX4720. To identify potential mechanisms of resistance to PLX4720 treatment and synergy with lapatinib treatment we performed a multi-platform functional genomics analysis to profile the genome as well as the transcriptional and proteomic responses of these cell lines to treatment with PLX4720. We found modest levels of resistance correlated with the zygosity of the BRAF V600E allele and RTK mutational status. Layered over base-line resistance was substantial upregulation of many ERBB pathway genes in response to BRaf inhibition, thus generating the vulnerability to combination with lapatinib. The transcriptional responses of ERBB pathway genes are associated with a number of transcription factors, including ETS2 and its associated cofactors that represent a convergent regulatory mechanism conferring synergistic drug susceptibility in the context of diverse mutational landscapes. 12 BRAF mutant melanomas and 4 melanomas with WT BRAF were exposed plx4720 treatment to evaluate their responses after 8 hours of treatment. 5 of the 12 BRAF mutant melanomas responses were also evaluated in response to the treatment of lapatinib alone, masitinib alone, the combination of lapatinib with plx4720, or the combination of masitinib with plx4720. All samples were run in at least triplicate.

Dataset Information

Antipermeability function of PEDF involves blockade of the MAP kinase/GSK/beta-catenin signaling pathway and uPAR expression.

Publications

Antipermeability function of PEDF involves blockade of the MAP kinase/GSK/beta-catenin signaling pathway and uPAR expression.

Similar Datasets

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