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Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts
PURPOSE: Oral wound healing requires gingival fibroblasts to respond to local growth factors. Epigenetic silencing through DNA methylation can potentially decrease the responsiveness of gingival fibroblasts to local growth factors. In this study, our aim was to determine whether the inhibition of DN...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Korean Academy of Periodontology
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5410554/ https://www.ncbi.nlm.nih.gov/pubmed/28462005 http://dx.doi.org/10.5051/jpis.2017.47.2.66 |
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author | Sufaru, Irina-Georgeta Beikircher, Gabriel Weinhaeusel, Andreas Gruber, Reinhard |
author_facet | Sufaru, Irina-Georgeta Beikircher, Gabriel Weinhaeusel, Andreas Gruber, Reinhard |
author_sort | Sufaru, Irina-Georgeta |
collection | PubMed |
description | PURPOSE: Oral wound healing requires gingival fibroblasts to respond to local growth factors. Epigenetic silencing through DNA methylation can potentially decrease the responsiveness of gingival fibroblasts to local growth factors. In this study, our aim was to determine whether the inhibition of DNA methylation sensitized gingival fibroblasts to transforming growth factor-β1 (TGF-β1). METHODS: Gingival fibroblasts were exposed to 5-aza-2'-deoxycytidine (5-aza), a clinically approved demethylating agent, before stimulation with TGF-β1. Gene expression changes were evaluated using quantitative polymerase chain reaction (PCR) analysis. DNA methylation was detected by methylation-sensitive restriction enzymes and PCR amplification. RESULTS: We found that 5-aza enhanced TGF-β1-induced interleukin-11 (IL11) expression in gingival fibroblasts 2.37-fold (P=0.008). 5-aza had no significant effects on the expression of proteoglycan 4 (PRG4) and NADPH oxidase 4 (NOX4). Consistent with this, 5-aza caused demethylation of the IL11 gene commonly next to a guanosine (CpG) island in gingival fibroblasts. The TGF-β type I receptor kinase inhibitor SB431542 impeded the changes in IL11 expression, indicating that the effects of 5-aza require TGF-β signaling. 5-aza moderately increased the expression of TGF-β type II receptor (1.40-fold; P=0.009), possibly enhancing the responsiveness of fibroblasts to TGF-β1. As part of the feedback response, 5-aza increased the expression of the DNA methyltransferases 1 (DNMT1) (P=0.005) and DNMT3B (P=0.002), which are enzymes responsible for gene methylation. CONCLUSIONS: These in vitro data suggest that the inhibition of DNA methylation by 5-aza supports TGF-β-induced IL11 expression in gingival fibroblasts. |
format | Online Article Text |
id | pubmed-5410554 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Korean Academy of Periodontology |
record_format | MEDLINE/PubMed |
spelling | pubmed-54105542017-05-01 Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts Sufaru, Irina-Georgeta Beikircher, Gabriel Weinhaeusel, Andreas Gruber, Reinhard J Periodontal Implant Sci Research Article PURPOSE: Oral wound healing requires gingival fibroblasts to respond to local growth factors. Epigenetic silencing through DNA methylation can potentially decrease the responsiveness of gingival fibroblasts to local growth factors. In this study, our aim was to determine whether the inhibition of DNA methylation sensitized gingival fibroblasts to transforming growth factor-β1 (TGF-β1). METHODS: Gingival fibroblasts were exposed to 5-aza-2'-deoxycytidine (5-aza), a clinically approved demethylating agent, before stimulation with TGF-β1. Gene expression changes were evaluated using quantitative polymerase chain reaction (PCR) analysis. DNA methylation was detected by methylation-sensitive restriction enzymes and PCR amplification. RESULTS: We found that 5-aza enhanced TGF-β1-induced interleukin-11 (IL11) expression in gingival fibroblasts 2.37-fold (P=0.008). 5-aza had no significant effects on the expression of proteoglycan 4 (PRG4) and NADPH oxidase 4 (NOX4). Consistent with this, 5-aza caused demethylation of the IL11 gene commonly next to a guanosine (CpG) island in gingival fibroblasts. The TGF-β type I receptor kinase inhibitor SB431542 impeded the changes in IL11 expression, indicating that the effects of 5-aza require TGF-β signaling. 5-aza moderately increased the expression of TGF-β type II receptor (1.40-fold; P=0.009), possibly enhancing the responsiveness of fibroblasts to TGF-β1. As part of the feedback response, 5-aza increased the expression of the DNA methyltransferases 1 (DNMT1) (P=0.005) and DNMT3B (P=0.002), which are enzymes responsible for gene methylation. CONCLUSIONS: These in vitro data suggest that the inhibition of DNA methylation by 5-aza supports TGF-β-induced IL11 expression in gingival fibroblasts. Korean Academy of Periodontology 2017-04 2017-04-29 /pmc/articles/PMC5410554/ /pubmed/28462005 http://dx.doi.org/10.5051/jpis.2017.47.2.66 Text en Copyright © 2017. Korean Academy of Periodontology https://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/). |
spellingShingle | Research Article Sufaru, Irina-Georgeta Beikircher, Gabriel Weinhaeusel, Andreas Gruber, Reinhard Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts |
title | Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts |
title_full | Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts |
title_fullStr | Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts |
title_full_unstemmed | Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts |
title_short | Inhibitors of DNA methylation support TGF-β1-induced IL11 expression in gingival fibroblasts |
title_sort | inhibitors of dna methylation support tgf-β1-induced il11 expression in gingival fibroblasts |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5410554/ https://www.ncbi.nlm.nih.gov/pubmed/28462005 http://dx.doi.org/10.5051/jpis.2017.47.2.66 |
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