Cargando…

17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells

Estrogens play a protective role in coronary artery disease. The mechanisms of action are still poorly understood, although a role for estrogens in stimulation of angiogenesis has been suggested. In several cell types, estrogens modulate the Notch pathway, which is involved in controlling angiogenes...

Descripción completa

Detalles Bibliográficos
Autores principales: Caliceti, Cristiana, Aquila, Giorgio, Pannella, Micaela, Morelli, Marco Bruno, Fortini, Cinzia, Pinton, Paolo, Bonora, Massimo, Hrelia, Silvana, Pannuti, Antonio, Miele, Lucio, Rizzo, Paola, Ferrari, Roberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3742772/
https://www.ncbi.nlm.nih.gov/pubmed/23967210
http://dx.doi.org/10.1371/journal.pone.0071440
_version_ 1782280409876267008
author Caliceti, Cristiana
Aquila, Giorgio
Pannella, Micaela
Morelli, Marco Bruno
Fortini, Cinzia
Pinton, Paolo
Bonora, Massimo
Hrelia, Silvana
Pannuti, Antonio
Miele, Lucio
Rizzo, Paola
Ferrari, Roberto
author_facet Caliceti, Cristiana
Aquila, Giorgio
Pannella, Micaela
Morelli, Marco Bruno
Fortini, Cinzia
Pinton, Paolo
Bonora, Massimo
Hrelia, Silvana
Pannuti, Antonio
Miele, Lucio
Rizzo, Paola
Ferrari, Roberto
author_sort Caliceti, Cristiana
collection PubMed
description Estrogens play a protective role in coronary artery disease. The mechanisms of action are still poorly understood, although a role for estrogens in stimulation of angiogenesis has been suggested. In several cell types, estrogens modulate the Notch pathway, which is involved in controlling angiogenesis downstream of vascular endothelial growth factor A (VEGF-A). The goal of our study was to establish whether estrogens modulate Notch activity in endothelial cells and the possible consequences on angiogenesis. Human umbilical vein endothelial cells (HUVECs) were treated with 17β-estradiol (E2) and the effects on Notch signalling were evaluated. E2 increased Notch1 processing as indicated by i) decreased levels of Notch1 transmembrane subunit ii) increased amount of Notch1 in nuclei iii) unaffected level of mRNA. Similarly, E2 increased the levels of the active form of Notch4 without altering Notch4 mRNA. Conversely, protein and mRNA levels of Notch2 were both reduced suggesting transcriptional repression of Notch2 by E2. Under conditions where Notch was activated by upregulation of Delta-like ligand 4 (Dll4) following VEGF-A treatment, E2 caused a further increase of the active form of Notch1, of the number of cells with nuclear Notch1 and of Hey2 mRNA. Estrogen receptor antagonist ICI 182.780 antagonized these effects suggesting that E2 modulation of Notch1 is mediated by estrogen receptors. E2 treatment abolished the increase in endothelial cells sprouting caused by Notch inhibition in a tube formation assay on 3D Matrigel and in mouse aortic ring explants. In conclusion, E2 affects several Notch pathway components in HUVECs, leading to an activation of the VEGF-A-Dll4-Notch1 axis and to a modulation of vascular branching when Notch signalling is inhibited. These results contribute to our understanding of the molecular mechanisms of cardiovascular protection exerted by estrogens by uncovering a novel role of E2 in the Notch signalling-mediated modulation of angiogenesis.
format Online
Article
Text
id pubmed-3742772
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-37427722013-08-21 17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells Caliceti, Cristiana Aquila, Giorgio Pannella, Micaela Morelli, Marco Bruno Fortini, Cinzia Pinton, Paolo Bonora, Massimo Hrelia, Silvana Pannuti, Antonio Miele, Lucio Rizzo, Paola Ferrari, Roberto PLoS One Research Article Estrogens play a protective role in coronary artery disease. The mechanisms of action are still poorly understood, although a role for estrogens in stimulation of angiogenesis has been suggested. In several cell types, estrogens modulate the Notch pathway, which is involved in controlling angiogenesis downstream of vascular endothelial growth factor A (VEGF-A). The goal of our study was to establish whether estrogens modulate Notch activity in endothelial cells and the possible consequences on angiogenesis. Human umbilical vein endothelial cells (HUVECs) were treated with 17β-estradiol (E2) and the effects on Notch signalling were evaluated. E2 increased Notch1 processing as indicated by i) decreased levels of Notch1 transmembrane subunit ii) increased amount of Notch1 in nuclei iii) unaffected level of mRNA. Similarly, E2 increased the levels of the active form of Notch4 without altering Notch4 mRNA. Conversely, protein and mRNA levels of Notch2 were both reduced suggesting transcriptional repression of Notch2 by E2. Under conditions where Notch was activated by upregulation of Delta-like ligand 4 (Dll4) following VEGF-A treatment, E2 caused a further increase of the active form of Notch1, of the number of cells with nuclear Notch1 and of Hey2 mRNA. Estrogen receptor antagonist ICI 182.780 antagonized these effects suggesting that E2 modulation of Notch1 is mediated by estrogen receptors. E2 treatment abolished the increase in endothelial cells sprouting caused by Notch inhibition in a tube formation assay on 3D Matrigel and in mouse aortic ring explants. In conclusion, E2 affects several Notch pathway components in HUVECs, leading to an activation of the VEGF-A-Dll4-Notch1 axis and to a modulation of vascular branching when Notch signalling is inhibited. These results contribute to our understanding of the molecular mechanisms of cardiovascular protection exerted by estrogens by uncovering a novel role of E2 in the Notch signalling-mediated modulation of angiogenesis. Public Library of Science 2013-08-13 /pmc/articles/PMC3742772/ /pubmed/23967210 http://dx.doi.org/10.1371/journal.pone.0071440 Text en © 2013 Caliceti et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Caliceti, Cristiana
Aquila, Giorgio
Pannella, Micaela
Morelli, Marco Bruno
Fortini, Cinzia
Pinton, Paolo
Bonora, Massimo
Hrelia, Silvana
Pannuti, Antonio
Miele, Lucio
Rizzo, Paola
Ferrari, Roberto
17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells
title 17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells
title_full 17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells
title_fullStr 17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells
title_full_unstemmed 17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells
title_short 17β-Estradiol Enhances Signalling Mediated by VEGF-A-Delta-Like Ligand 4-Notch1 Axis in Human Endothelial Cells
title_sort 17β-estradiol enhances signalling mediated by vegf-a-delta-like ligand 4-notch1 axis in human endothelial cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3742772/
https://www.ncbi.nlm.nih.gov/pubmed/23967210
http://dx.doi.org/10.1371/journal.pone.0071440
work_keys_str_mv AT caliceticristiana 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT aquilagiorgio 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT pannellamicaela 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT morellimarcobruno 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT fortinicinzia 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT pintonpaolo 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT bonoramassimo 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT hreliasilvana 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT pannutiantonio 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT mielelucio 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT rizzopaola 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells
AT ferrariroberto 17bestradiolenhancessignallingmediatedbyvegfadeltalikeligand4notch1axisinhumanendothelialcells