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MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells

A rationally selected combination of small-molecule chemicals can affect cell plasticity and fate, suggesting an open chemistry way to manipulate cells to achieve a specific goal. Here we for the first time demonstrate that a combination of vitamin C (Vc) and PD0325901 can achieve about 90% erasure...

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Autores principales: Li, Cuiping, Liu, Baodong, Zhong, Shangwei, Wang, Hailin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5129966/
https://www.ncbi.nlm.nih.gov/pubmed/27213595
http://dx.doi.org/10.18632/oncotarget.9452
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author Li, Cuiping
Liu, Baodong
Zhong, Shangwei
Wang, Hailin
author_facet Li, Cuiping
Liu, Baodong
Zhong, Shangwei
Wang, Hailin
author_sort Li, Cuiping
collection PubMed
description A rationally selected combination of small-molecule chemicals can affect cell plasticity and fate, suggesting an open chemistry way to manipulate cells to achieve a specific goal. Here we for the first time demonstrate that a combination of vitamin C (Vc) and PD0325901 can achieve about 90% erasure of 5-methylcytosine (5mC) within 5 days (decreasing from 3.2 to ~ 0.3 5mC per 100 C) in mouse embryonic stem cells (ESCs). The hypomethylated level is comparable to that of gonadal primordial germ cells (PGCs), whose pluripotency is closely associated with the global DNA hypomethylation. In contrast, Vc or PD0325901 alone only induces a moderately reduced level of global DNA methylation. Our mechanistic study suggested that PD0325901 elevated expression of Prdm14, which repressed de novo methyltransferase Dnmt3b and its cofactor Dnmt3l at levels of protein, via the mode to eliminate 5mC from de novo synthesis. By further addition of Vc, the oxidation of 5mC as catalyzed by Tet1/Tet2 dioxygenases was significantly increased as manifested by the elevated level of 5-hydroxymethylcytosine. However, by the depletion of Tet1/Tet2, Vc failed to enhance PD0325901-stimulated hypomethylation of ESCs’ genomic DNA. Furthermore, mouse ESCs in Vc/PD0325901-supplemented medium show great morphology and pluripotency. Therefore, we demonstrate a novel and synergistic chemical approach for promoting hypomethylation and sustaining pluripotency of ESCs.
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spelling pubmed-51299662016-12-11 MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells Li, Cuiping Liu, Baodong Zhong, Shangwei Wang, Hailin Oncotarget Research Paper A rationally selected combination of small-molecule chemicals can affect cell plasticity and fate, suggesting an open chemistry way to manipulate cells to achieve a specific goal. Here we for the first time demonstrate that a combination of vitamin C (Vc) and PD0325901 can achieve about 90% erasure of 5-methylcytosine (5mC) within 5 days (decreasing from 3.2 to ~ 0.3 5mC per 100 C) in mouse embryonic stem cells (ESCs). The hypomethylated level is comparable to that of gonadal primordial germ cells (PGCs), whose pluripotency is closely associated with the global DNA hypomethylation. In contrast, Vc or PD0325901 alone only induces a moderately reduced level of global DNA methylation. Our mechanistic study suggested that PD0325901 elevated expression of Prdm14, which repressed de novo methyltransferase Dnmt3b and its cofactor Dnmt3l at levels of protein, via the mode to eliminate 5mC from de novo synthesis. By further addition of Vc, the oxidation of 5mC as catalyzed by Tet1/Tet2 dioxygenases was significantly increased as manifested by the elevated level of 5-hydroxymethylcytosine. However, by the depletion of Tet1/Tet2, Vc failed to enhance PD0325901-stimulated hypomethylation of ESCs’ genomic DNA. Furthermore, mouse ESCs in Vc/PD0325901-supplemented medium show great morphology and pluripotency. Therefore, we demonstrate a novel and synergistic chemical approach for promoting hypomethylation and sustaining pluripotency of ESCs. Impact Journals LLC 2016-05-18 /pmc/articles/PMC5129966/ /pubmed/27213595 http://dx.doi.org/10.18632/oncotarget.9452 Text en Copyright: © 2016 Li et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Li, Cuiping
Liu, Baodong
Zhong, Shangwei
Wang, Hailin
MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells
title MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells
title_full MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells
title_fullStr MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells
title_full_unstemmed MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells
title_short MEK inhibitor PD0325901 and vitamin C synergistically induce hypomethylation of mouse embryonic stem cells
title_sort mek inhibitor pd0325901 and vitamin c synergistically induce hypomethylation of mouse embryonic stem cells
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5129966/
https://www.ncbi.nlm.nih.gov/pubmed/27213595
http://dx.doi.org/10.18632/oncotarget.9452
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