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DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction

OBJECTIVE: Cellular repressor of E1A-stimulated genes (CREG), a vasculoprotective molecule, is significantly downregulated in atherosclerotic vessels through unclear mechanisms. While epigenetic regulation is involved in atherosclerosis development, it is not known if the CREG gene is epigenetically...

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Autores principales: Liu, Yanxia, Tian, Xiaoxiang, Liu, Shan, Liu, Dan, Li, Yang, Liu, Meili, Zhang, Xiaolin, Yan, Chenghui, Han, Yaling
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264464/
https://www.ncbi.nlm.nih.gov/pubmed/32067910
http://dx.doi.org/10.1016/j.redox.2020.101444
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author Liu, Yanxia
Tian, Xiaoxiang
Liu, Shan
Liu, Dan
Li, Yang
Liu, Meili
Zhang, Xiaolin
Yan, Chenghui
Han, Yaling
author_facet Liu, Yanxia
Tian, Xiaoxiang
Liu, Shan
Liu, Dan
Li, Yang
Liu, Meili
Zhang, Xiaolin
Yan, Chenghui
Han, Yaling
author_sort Liu, Yanxia
collection PubMed
description OBJECTIVE: Cellular repressor of E1A-stimulated genes (CREG), a vasculoprotective molecule, is significantly downregulated in atherosclerotic vessels through unclear mechanisms. While epigenetic regulation is involved in atherosclerosis development, it is not known if the CREG gene is epigenetically regulated. The aim of this study was to assess the potential role of CREG methylation in contributing to atherosclerosis. APPROACH AND RESULTS: Overexpression of DNA methyltransferase (DNMT)3B significantly inhibited CREG expression in human umbilical vein endothelial cells (HUVECs) and human coronary aortic endothelial cells (HCAECs).Conversely, inhibition of DNA methylation with 5-aza-2′-deoxycytidine (5-aza-dC) dose-dependently increased CREG expression. A CREG promoter analysis identified +168 to +255 bp as a key regulatory region and the CG site at +201/+202 bp as a key methylation site. The transcription factor GR-α could bind to the +201/+202 bp CG site promoting CREG transcription, a process significantly inhibited by DNMT3B overexpression. Treatment of cells with oxidized low-density lipoprotein (ox-LDL), a critical atherosclerogenic factor, significantly increased DNMT3B expression, increasing CREG promotor methylation, blocking GR-α binding, and inhibiting CREG expression. Consistently, CG sites in the CREG promoter fragment were hyper-methylated in human atherosclerotic arteries, and CREG expression was significantly reduced. A negative correlation between DNMT3B and CREG expression levels was observed in human atherosclerotic arteries. Finally, Ox-LDL-induced endothelium dysfunction was significantly attenuated by both 5-aza-dC and an anti-oxidative molecular N-acetylcysteine (NAC) administration through rescue the expression of CREG and activation of the p-eNOS/NO pathway. CONCLUSIONS: Our study provides the first direct evidence that DNMT3B-mediated CREG gene hypermethylation is a novel mechanism that contributes to endothelial dysfunction and atherosclerosis development. Blocking CREG methylation may represent a novel therapeutic approach to treat ox-LDL-induced atherosclerosis.
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spelling pubmed-72644642020-06-05 DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction Liu, Yanxia Tian, Xiaoxiang Liu, Shan Liu, Dan Li, Yang Liu, Meili Zhang, Xiaolin Yan, Chenghui Han, Yaling Redox Biol Research Paper OBJECTIVE: Cellular repressor of E1A-stimulated genes (CREG), a vasculoprotective molecule, is significantly downregulated in atherosclerotic vessels through unclear mechanisms. While epigenetic regulation is involved in atherosclerosis development, it is not known if the CREG gene is epigenetically regulated. The aim of this study was to assess the potential role of CREG methylation in contributing to atherosclerosis. APPROACH AND RESULTS: Overexpression of DNA methyltransferase (DNMT)3B significantly inhibited CREG expression in human umbilical vein endothelial cells (HUVECs) and human coronary aortic endothelial cells (HCAECs).Conversely, inhibition of DNA methylation with 5-aza-2′-deoxycytidine (5-aza-dC) dose-dependently increased CREG expression. A CREG promoter analysis identified +168 to +255 bp as a key regulatory region and the CG site at +201/+202 bp as a key methylation site. The transcription factor GR-α could bind to the +201/+202 bp CG site promoting CREG transcription, a process significantly inhibited by DNMT3B overexpression. Treatment of cells with oxidized low-density lipoprotein (ox-LDL), a critical atherosclerogenic factor, significantly increased DNMT3B expression, increasing CREG promotor methylation, blocking GR-α binding, and inhibiting CREG expression. Consistently, CG sites in the CREG promoter fragment were hyper-methylated in human atherosclerotic arteries, and CREG expression was significantly reduced. A negative correlation between DNMT3B and CREG expression levels was observed in human atherosclerotic arteries. Finally, Ox-LDL-induced endothelium dysfunction was significantly attenuated by both 5-aza-dC and an anti-oxidative molecular N-acetylcysteine (NAC) administration through rescue the expression of CREG and activation of the p-eNOS/NO pathway. CONCLUSIONS: Our study provides the first direct evidence that DNMT3B-mediated CREG gene hypermethylation is a novel mechanism that contributes to endothelial dysfunction and atherosclerosis development. Blocking CREG methylation may represent a novel therapeutic approach to treat ox-LDL-induced atherosclerosis. Elsevier 2020-01-31 /pmc/articles/PMC7264464/ /pubmed/32067910 http://dx.doi.org/10.1016/j.redox.2020.101444 Text en © 2020 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Paper
Liu, Yanxia
Tian, Xiaoxiang
Liu, Shan
Liu, Dan
Li, Yang
Liu, Meili
Zhang, Xiaolin
Yan, Chenghui
Han, Yaling
DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction
title DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction
title_full DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction
title_fullStr DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction
title_full_unstemmed DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction
title_short DNA hypermethylation: A novel mechanism of CREG gene suppression and atherosclerogenic endothelial dysfunction
title_sort dna hypermethylation: a novel mechanism of creg gene suppression and atherosclerogenic endothelial dysfunction
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264464/
https://www.ncbi.nlm.nih.gov/pubmed/32067910
http://dx.doi.org/10.1016/j.redox.2020.101444
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