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Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing

Long noncoding RNAs (lncRNAs) are non-protein coding RNAs regulating gene expression. Although for some lncRNAs a relevant role in hypoxic endothelium has been shown, the regulation and function of lncRNAs is still largely unknown in the vascular physio-pathology. Taking advantage of next-generation...

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Autores principales: Voellenkle, C., Garcia-Manteiga, J. M., Pedrotti, S., Perfetti, A., De Toma, I., Da Silva, D., Maimone, B., Greco, S., Fasanaro, P., Creo, P., Zaccagnini, G., Gaetano, C., Martelli, F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4827084/
https://www.ncbi.nlm.nih.gov/pubmed/27063004
http://dx.doi.org/10.1038/srep24141
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author Voellenkle, C.
Garcia-Manteiga, J. M.
Pedrotti, S.
Perfetti, A.
De Toma, I.
Da Silva, D.
Maimone, B.
Greco, S.
Fasanaro, P.
Creo, P.
Zaccagnini, G.
Gaetano, C.
Martelli, F.
author_facet Voellenkle, C.
Garcia-Manteiga, J. M.
Pedrotti, S.
Perfetti, A.
De Toma, I.
Da Silva, D.
Maimone, B.
Greco, S.
Fasanaro, P.
Creo, P.
Zaccagnini, G.
Gaetano, C.
Martelli, F.
author_sort Voellenkle, C.
collection PubMed
description Long noncoding RNAs (lncRNAs) are non-protein coding RNAs regulating gene expression. Although for some lncRNAs a relevant role in hypoxic endothelium has been shown, the regulation and function of lncRNAs is still largely unknown in the vascular physio-pathology. Taking advantage of next-generation sequencing techniques, transcriptomic changes induced by endothelial cell exposure to hypoxia were investigated. Paired-end sequencing of polyadenylated RNA derived from human umbilical vein endothelial cells (HUVECs) exposed to 1% O(2) or normoxia was performed. Bioinformatics analysis identified ≈2000 differentially expressed genes, including 122 lncRNAs. Extensive validation was performed by both microarray and qPCR. Among the validated lncRNAs, H19, MIR210HG, MEG9, MALAT1 and MIR22HG were also induced in a mouse model of hindlimb ischemia. To test the functional relevance of lncRNAs in endothelial cells, knockdown of H19 expression was performed. H19 inhibition decreased HUVEC growth, inducing their accumulation in G1 phase of the cell cycle; accordingly, p21 (CDKN1A) expression was increased. Additionally, H19 knockdown also diminished HUVEC ability to form capillary like structures when plated on matrigel. In conclusion, a high-confidence signature of lncRNAs modulated by hypoxia in HUVEC was identified and a significant impact of H19 lncRNA was shown.
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spelling pubmed-48270842016-04-19 Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing Voellenkle, C. Garcia-Manteiga, J. M. Pedrotti, S. Perfetti, A. De Toma, I. Da Silva, D. Maimone, B. Greco, S. Fasanaro, P. Creo, P. Zaccagnini, G. Gaetano, C. Martelli, F. Sci Rep Article Long noncoding RNAs (lncRNAs) are non-protein coding RNAs regulating gene expression. Although for some lncRNAs a relevant role in hypoxic endothelium has been shown, the regulation and function of lncRNAs is still largely unknown in the vascular physio-pathology. Taking advantage of next-generation sequencing techniques, transcriptomic changes induced by endothelial cell exposure to hypoxia were investigated. Paired-end sequencing of polyadenylated RNA derived from human umbilical vein endothelial cells (HUVECs) exposed to 1% O(2) or normoxia was performed. Bioinformatics analysis identified ≈2000 differentially expressed genes, including 122 lncRNAs. Extensive validation was performed by both microarray and qPCR. Among the validated lncRNAs, H19, MIR210HG, MEG9, MALAT1 and MIR22HG were also induced in a mouse model of hindlimb ischemia. To test the functional relevance of lncRNAs in endothelial cells, knockdown of H19 expression was performed. H19 inhibition decreased HUVEC growth, inducing their accumulation in G1 phase of the cell cycle; accordingly, p21 (CDKN1A) expression was increased. Additionally, H19 knockdown also diminished HUVEC ability to form capillary like structures when plated on matrigel. In conclusion, a high-confidence signature of lncRNAs modulated by hypoxia in HUVEC was identified and a significant impact of H19 lncRNA was shown. Nature Publishing Group 2016-04-11 /pmc/articles/PMC4827084/ /pubmed/27063004 http://dx.doi.org/10.1038/srep24141 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Voellenkle, C.
Garcia-Manteiga, J. M.
Pedrotti, S.
Perfetti, A.
De Toma, I.
Da Silva, D.
Maimone, B.
Greco, S.
Fasanaro, P.
Creo, P.
Zaccagnini, G.
Gaetano, C.
Martelli, F.
Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing
title Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing
title_full Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing
title_fullStr Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing
title_full_unstemmed Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing
title_short Implication of Long noncoding RNAs in the endothelial cell response to hypoxia revealed by RNA-sequencing
title_sort implication of long noncoding rnas in the endothelial cell response to hypoxia revealed by rna-sequencing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4827084/
https://www.ncbi.nlm.nih.gov/pubmed/27063004
http://dx.doi.org/10.1038/srep24141
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