RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0207873 http://hdl.handle.net/11449/185146 |
Resumo: | Human bone marrow-derived mesenchymal stem cells (hBMSCs) are important for tissue regeneration but their epigenetic regulation is not well understood. Here we investigate the ability of a non-nucleoside DNA methylation inhibitor, RG108 to induce epigenetic changes at both global and gene-specific levels in order to enhance mesenchymal cell markers, in hBMSCs. hBMSCs were treated with complete culture medium, 50 mu M RG108 and DMSO for three days and subjected to viability and apoptosis assays, global and gene-specific methylation/hydroxymethylation, transcript levels' analysis of epigenetic machinery enzymes and multipotency markers, protein activities of DNMTs and TETs, immunofluorescence staining and western blot analysis for NANOG and OCT4 and flow cytometry for CD105. The RG108, when used at 50 mu M, did not affect the viability, apoptosis and proliferation rates of hBMSCs or hydroxymethylation global levels while leading to 75% decrease in DNMTs activity and 42% loss of global DNA methylation levels. In addition, DNMT1 was significantly downregulated while TET1 was upregulated, potentially contributing to the substantial loss of methylation observed. Most importantly, the mesenchymal cell markers CD105, NANOG and OCT4 were upregulated being NANOG and OCT4 epigenetically modulated by RG108, at their gene promoters. We propose that RG108 could be used for epigenetic modulation, promoting epigenetic activation of NANOG and OCT4, without affecting the viability of hBMSCs. DMSO can be considered a modulator of epigenetic machinery enzymes, although with milder effect compared to RG108. |
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2946 |
spelling |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activationHuman bone marrow-derived mesenchymal stem cells (hBMSCs) are important for tissue regeneration but their epigenetic regulation is not well understood. Here we investigate the ability of a non-nucleoside DNA methylation inhibitor, RG108 to induce epigenetic changes at both global and gene-specific levels in order to enhance mesenchymal cell markers, in hBMSCs. hBMSCs were treated with complete culture medium, 50 mu M RG108 and DMSO for three days and subjected to viability and apoptosis assays, global and gene-specific methylation/hydroxymethylation, transcript levels' analysis of epigenetic machinery enzymes and multipotency markers, protein activities of DNMTs and TETs, immunofluorescence staining and western blot analysis for NANOG and OCT4 and flow cytometry for CD105. The RG108, when used at 50 mu M, did not affect the viability, apoptosis and proliferation rates of hBMSCs or hydroxymethylation global levels while leading to 75% decrease in DNMTs activity and 42% loss of global DNA methylation levels. In addition, DNMT1 was significantly downregulated while TET1 was upregulated, potentially contributing to the substantial loss of methylation observed. Most importantly, the mesenchymal cell markers CD105, NANOG and OCT4 were upregulated being NANOG and OCT4 epigenetically modulated by RG108, at their gene promoters. We propose that RG108 could be used for epigenetic modulation, promoting epigenetic activation of NANOG and OCT4, without affecting the viability of hBMSCs. DMSO can be considered a modulator of epigenetic machinery enzymes, although with milder effect compared to RG108.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FP7 Framework Marie Curie Actions Career Integration Grant (methDRE)Univ Estadual Campinas, Piracicaba Dent Sch, Dept Prosthodont & Periodont, Piracicaba, SP, BrazilUniv Birmingham, Sch Canc Sci, Sch Dent, Birmingham, W Midlands, EnglandSao Paulo State Univ, Biosci Inst, Dept Chem & Biochem, Botucatu, SP, BrazilUniv Paulista, Sch Dent, Div Epigenet, Hlth Sci Inst, Sao Paulo, BrazilSao Paulo State Univ, Biosci Inst, Dept Chem & Biochem, Botucatu, SP, BrazilFAPESP: 2013/09650-8FAPESP: 2015/02160-0Public Library ScienceUniversidade Estadual de Campinas (UNICAMP)Univ BirminghamUniversidade Estadual Paulista (Unesp)Univ PaulistaAssis, Rahyza I. F.Wiench, MalgorzataSilverio, Karina G.Silva, Rodrigo A. da [UNESP]Feltran, Georgia da Silva [UNESP]Sallum, Enilson A.Casati, Marcio Z.Nociti, Francisco H.Andia, Denise C.2019-10-04T12:33:03Z2019-10-04T12:33:03Z2018-12-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article20http://dx.doi.org/10.1371/journal.pone.0207873Plos One. San Francisco: Public Library Science, v. 13, n. 12, 20 p., 2018.1932-6203http://hdl.handle.net/11449/18514610.1371/journal.pone.0207873WOS:000451886500013Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPlos Oneinfo:eu-repo/semantics/openAccess2021-10-23T19:02:04Zoai:repositorio.unesp.br:11449/185146Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T18:11:51.814965Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
title |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
spellingShingle |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation Assis, Rahyza I. F. |
title_short |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
title_full |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
title_fullStr |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
title_full_unstemmed |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
title_sort |
RG108 increases NANOG and OCT4 in bone marrow-derived mesenchymal cells through global changes in DNA modifications and epigenetic activation |
author |
Assis, Rahyza I. F. |
author_facet |
Assis, Rahyza I. F. Wiench, Malgorzata Silverio, Karina G. Silva, Rodrigo A. da [UNESP] Feltran, Georgia da Silva [UNESP] Sallum, Enilson A. Casati, Marcio Z. Nociti, Francisco H. Andia, Denise C. |
author_role |
author |
author2 |
Wiench, Malgorzata Silverio, Karina G. Silva, Rodrigo A. da [UNESP] Feltran, Georgia da Silva [UNESP] Sallum, Enilson A. Casati, Marcio Z. Nociti, Francisco H. Andia, Denise C. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual de Campinas (UNICAMP) Univ Birmingham Universidade Estadual Paulista (Unesp) Univ Paulista |
dc.contributor.author.fl_str_mv |
Assis, Rahyza I. F. Wiench, Malgorzata Silverio, Karina G. Silva, Rodrigo A. da [UNESP] Feltran, Georgia da Silva [UNESP] Sallum, Enilson A. Casati, Marcio Z. Nociti, Francisco H. Andia, Denise C. |
description |
Human bone marrow-derived mesenchymal stem cells (hBMSCs) are important for tissue regeneration but their epigenetic regulation is not well understood. Here we investigate the ability of a non-nucleoside DNA methylation inhibitor, RG108 to induce epigenetic changes at both global and gene-specific levels in order to enhance mesenchymal cell markers, in hBMSCs. hBMSCs were treated with complete culture medium, 50 mu M RG108 and DMSO for three days and subjected to viability and apoptosis assays, global and gene-specific methylation/hydroxymethylation, transcript levels' analysis of epigenetic machinery enzymes and multipotency markers, protein activities of DNMTs and TETs, immunofluorescence staining and western blot analysis for NANOG and OCT4 and flow cytometry for CD105. The RG108, when used at 50 mu M, did not affect the viability, apoptosis and proliferation rates of hBMSCs or hydroxymethylation global levels while leading to 75% decrease in DNMTs activity and 42% loss of global DNA methylation levels. In addition, DNMT1 was significantly downregulated while TET1 was upregulated, potentially contributing to the substantial loss of methylation observed. Most importantly, the mesenchymal cell markers CD105, NANOG and OCT4 were upregulated being NANOG and OCT4 epigenetically modulated by RG108, at their gene promoters. We propose that RG108 could be used for epigenetic modulation, promoting epigenetic activation of NANOG and OCT4, without affecting the viability of hBMSCs. DMSO can be considered a modulator of epigenetic machinery enzymes, although with milder effect compared to RG108. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-03 2019-10-04T12:33:03Z 2019-10-04T12:33:03Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1371/journal.pone.0207873 Plos One. San Francisco: Public Library Science, v. 13, n. 12, 20 p., 2018. 1932-6203 http://hdl.handle.net/11449/185146 10.1371/journal.pone.0207873 WOS:000451886500013 |
url |
http://dx.doi.org/10.1371/journal.pone.0207873 http://hdl.handle.net/11449/185146 |
identifier_str_mv |
Plos One. San Francisco: Public Library Science, v. 13, n. 12, 20 p., 2018. 1932-6203 10.1371/journal.pone.0207873 WOS:000451886500013 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Plos One |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
20 |
dc.publisher.none.fl_str_mv |
Public Library Science |
publisher.none.fl_str_mv |
Public Library Science |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1808128907032920064 |