Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration
Background/Aim: Endothelial cell migration is required for physiological angiogenesis, but also contributes to various pathological conditions, including tumour vascularization. The mRNA expression of PP1cβ, the beta isoform of the catalytic PP1 subunit, was shown to be upregulated in chronic hypoxi...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Cell Physiol Biochem Press GmbH & Co KG
2015-05-01
|
Series: | Cellular Physiology and Biochemistry |
Subjects: | |
Online Access: | http://www.karger.com/Article/FullText/430257 |
id |
doaj-4b5064306ccc4baa94571ec9df67352f |
---|---|
record_format |
Article |
spelling |
doaj-4b5064306ccc4baa94571ec9df67352f2020-11-25T01:38:00ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782015-05-0136138439410.1159/000430257430257Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell MigrationDominga IacobazziIndira GaraevaAmbra AlbertarioMyriam CherifGianni D. AngeliniMassimo CaputoMohamed T. GhorbelBackground/Aim: Endothelial cell migration is required for physiological angiogenesis, but also contributes to various pathological conditions, including tumour vascularization. The mRNA expression of PP1cβ, the beta isoform of the catalytic PP1 subunit, was shown to be upregulated in chronic hypoxia. Since hypoxia is a major regulator of angiogenesis, the potential role of PP1cβ in angiogenesis was investigated. Methods: We examined PP1cβ protein level in pediatric heart following chronic hypoxia and found PP1cβ upregulation in cyanotic compared with acyanotic myocardium. By treating HUVEC cells with hypoxia mimicking agent, PP1cβ protein level increased with maximum at 8 hours. The effect of PP1cβ pharmacological inhibition, knockdown and overexpression, on endothelial cell migration and morphogenesis, was examined using in vitro wound healing scratch assay and endothelial tube formation assay. The PP1cβ knockdown effects on F-actin reorganization (phalloidin staining), focal adhesion formation (vinculin) and focal adhesion kinases (FAK) activation, were evaluated by immunocytochemical staining and immunoblotting with specific antibodies. Results: PP1cβ knockdown significantly reduces endothelial cell migration, but does not have any significant effect on endothelial tube formation. Endothelial cell migration in the knockdown group is restored to the control level upon consecutive transfection with PP1cβ cDNA. PP1cβ overexpression does not significantly affect endothelial cell migration. Furthermore, PP1cβ knockdown induces profound cytoskeletal reorganization, loss of focal adhesion sites and impairment of focal adhesion kinases (FAK) activation. Conclusions: PP1cβ is regulator of endothelial cell migration, which is critical in the angiogenic process. PP1cβ inhibition reduces endothelial cell migration through focal adhesion turnover and actin polymerization pathways.http://www.karger.com/Article/FullText/430257Protein phosphatase 1Endothelial cell migrationAngiogenesisHypoxia |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Dominga Iacobazzi Indira Garaeva Ambra Albertario Myriam Cherif Gianni D. Angelini Massimo Caputo Mohamed T. Ghorbel |
spellingShingle |
Dominga Iacobazzi Indira Garaeva Ambra Albertario Myriam Cherif Gianni D. Angelini Massimo Caputo Mohamed T. Ghorbel Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration Cellular Physiology and Biochemistry Protein phosphatase 1 Endothelial cell migration Angiogenesis Hypoxia |
author_facet |
Dominga Iacobazzi Indira Garaeva Ambra Albertario Myriam Cherif Gianni D. Angelini Massimo Caputo Mohamed T. Ghorbel |
author_sort |
Dominga Iacobazzi |
title |
Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration |
title_short |
Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration |
title_full |
Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration |
title_fullStr |
Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration |
title_full_unstemmed |
Protein Phosphatase 1 Beta is Modulated by Chronic Hypoxia and Involved in the Angiogenic Endothelial Cell Migration |
title_sort |
protein phosphatase 1 beta is modulated by chronic hypoxia and involved in the angiogenic endothelial cell migration |
publisher |
Cell Physiol Biochem Press GmbH & Co KG |
series |
Cellular Physiology and Biochemistry |
issn |
1015-8987 1421-9778 |
publishDate |
2015-05-01 |
description |
Background/Aim: Endothelial cell migration is required for physiological angiogenesis, but also contributes to various pathological conditions, including tumour vascularization. The mRNA expression of PP1cβ, the beta isoform of the catalytic PP1 subunit, was shown to be upregulated in chronic hypoxia. Since hypoxia is a major regulator of angiogenesis, the potential role of PP1cβ in angiogenesis was investigated. Methods: We examined PP1cβ protein level in pediatric heart following chronic hypoxia and found PP1cβ upregulation in cyanotic compared with acyanotic myocardium. By treating HUVEC cells with hypoxia mimicking agent, PP1cβ protein level increased with maximum at 8 hours. The effect of PP1cβ pharmacological inhibition, knockdown and overexpression, on endothelial cell migration and morphogenesis, was examined using in vitro wound healing scratch assay and endothelial tube formation assay. The PP1cβ knockdown effects on F-actin reorganization (phalloidin staining), focal adhesion formation (vinculin) and focal adhesion kinases (FAK) activation, were evaluated by immunocytochemical staining and immunoblotting with specific antibodies. Results: PP1cβ knockdown significantly reduces endothelial cell migration, but does not have any significant effect on endothelial tube formation. Endothelial cell migration in the knockdown group is restored to the control level upon consecutive transfection with PP1cβ cDNA. PP1cβ overexpression does not significantly affect endothelial cell migration. Furthermore, PP1cβ knockdown induces profound cytoskeletal reorganization, loss of focal adhesion sites and impairment of focal adhesion kinases (FAK) activation. Conclusions: PP1cβ is regulator of endothelial cell migration, which is critical in the angiogenic process. PP1cβ inhibition reduces endothelial cell migration through focal adhesion turnover and actin polymerization pathways. |
topic |
Protein phosphatase 1 Endothelial cell migration Angiogenesis Hypoxia |
url |
http://www.karger.com/Article/FullText/430257 |
work_keys_str_mv |
AT domingaiacobazzi proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration AT indiragaraeva proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration AT ambraalbertario proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration AT myriamcherif proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration AT giannidangelini proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration AT massimocaputo proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration AT mohamedtghorbel proteinphosphatase1betaismodulatedbychronichypoxiaandinvolvedintheangiogenicendothelialcellmigration |
_version_ |
1725055798247161856 |