Regulation of VEGF-activated signalling by the plasma membrane calcium ATPase 4 in endothelial cells
type: CreativeWork
description: INTRODUCTION: Angiogenesis, the formation of new blood vessels from pre-existing ones. It is a tightly regulated processes involving pro- and anti-angiogenic molecules. Deregulation of this process is associated with aberrant blood vessel formation (excessive or insufficient) in several human pathologies. Among the many pro-angiogenic factors promoting angiogenesis, the vascular endothelial growth factor (VEGF) has been characterised as a major regulator of both physiological and pathological angiogenesis. Therefore, the characterisation of the molecular mechanisms that regulate VEGF-induced angiogenesis is essential to develop therapeutic strategies that correct abnormal angiogenesis. In this sense, our group has previously reported a negative role for the Plasma Membrane Calcium ATPase 4 (PMCA4) protein in endothelial cells, acting via inhibition of the pro-angiogenic calcineurin/NFAT signalling pathway. However, we hypothesise that other intracellular pathways might be regulated by PMCA4 in endothelial cells during VEGF stimulation of angiogenesis. METHODS: To identify PMCA4 regulated pro-angiogenic signalling pathways, we have screened gene arrays related to Notch signalling or extracellular matrix-Cell Adhesion Molecule (ECM-CAM) pathway using RNA isolated from PMCA4-silenced (or control) HUVEC. Changes in gene expression after PMCA4 knockdown have been further validated by TaqMan-based qPCR in HUVEC or HDMEC. RNA levels of PMCA4 in aging HUVEC were analysed by TaqMan qPCR using RNA isolated from HUVEC cultured from different passages (from 3 to 15 passages). RESULTS: siRNA-mediated PMCA4 knockdown led to increased expression of Notch ligand DLL1 and Notch target gene Hey1 in VEGF-stimulated HUVEC. Expression of the transcription factor c-Fos was also elevated after PMCA4 knockdown in HUVEC stimulated with VEGF for 1h. Analysis of a gene array containing genes encoding extracellular matrix and cell adhesion molecules revealed that PMCA4 silencing alters the bas…
abstract: INTRODUCTION: Angiogenesis, the formation of new blood vessels from pre-existing ones. It is a tightly regulated processes involving pro- and anti-angiogenic molecules. Deregulation of this process is associated with aberrant blood vessel formation (excessive or insufficient) in several human pathologies. Among the many pro-angiogenic factors promoting angiogenesis, the vascular endothelial growth factor (VEGF) has been characterised as a major regulator of both physiological and pathological angiogenesis. Therefore, the characterisation of the molecular mechanisms that regulate VEGF-induced angiogenesis is essential to develop therapeutic strategies that correct abnormal angiogenesis. In this sense, our group has previously reported a negative role for the Plasma Membrane Calcium ATPase 4 (PMCA4) protein in endothelial cells, acting via inhibition of the pro-angiogenic calcineurin/NFAT signalling pathway. However, we hypothesise that other intracellular pathways might be regulated by PMCA4 in endothelial cells during VEGF stimulation of angiogenesis. METHODS: To identify PMCA4 regulated pro-angiogenic signalling pathways, we have screened gene arrays related to Notch signalling or extracellular matrix-Cell Adhesion Molecule (ECM-CAM) pathway using RNA isolated from PMCA4-silenced (or control) HUVEC. Changes in gene expression after PMCA4 knockdown have been further validated by TaqMan-based qPCR in HUVEC or HDMEC. RNA levels of PMCA4 in aging HUVEC were analysed by TaqMan qPCR using RNA isolated from HUVEC cultured from different passages (from 3 to 15 passages). RESULTS: siRNA-mediated PMCA4 knockdown led to increased expression of Notch ligand DLL1 and Notch target gene Hey1 in VEGF-stimulated HUVEC. Expression of the transcription factor c-Fos was also elevated after PMCA4 knockdown in HUVEC stimulated with VEGF for 1h. Analysis of a gene array containing genes encoding extracellular matrix and cell adhesion molecules revealed that PMCA4 silencing alters the bas…
author: Immanuel, Reshma Naomi Ranjit
keywords: VEGF signalling, notch signalling, ECM-CAM signalling, regulation of VEGF-activated signalling, HUVEC, HDMEC, PMCA4 gene silencing, DLL1 upregulation, cFOS upregulation, ADAMTS-1 upregulation
license: https://wlv.dspace7.openrepository.com/bitstreams/87c6fa0b-5a7f-46f0-aaf6-f6308ea842f4/download
date published: 2021-01
date modified: 2026-07-24T06:08:54Z
provider: University of Wolverhampton
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Wolverhampton