Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/328143
Type: Outros documentos
Title: Effects Of Epigenetic Modulator Drugs On Dna Methylation Of Hela Cells
Effects of epigenetic modulator drugs on DNA methylation of HeLa cells
Author: Veronezi, Giovana Maria Breda
Felisbino, Marina Barreto
Gatti, Maria Silvia Viccari
Vidal, Benedicto de Campos
Mello, Maria Luiza Silveira
Abstract: Valproic acid (VPA) is an important epigenetic drug that works as a histone deacetylase (HDAC) inhibitor, inducing histone hyperacetylation and chromatin unpackaging in several cell types. Currently, DNA demethylation is also being considered part of the VPA action, as shown in HEK293 cells and rat brain cells. This process may occur in a replication-independent active way, possibly with TET proteins participation and 5-hydroxymethylcytosine (5hmC) formation. Recent studies have also detected a 5hmC increase in different cell types after treatment with the well-established DNA methyltransferase (DNMT) inhibitor 5-aza-2'-deoxycytidine (5-aza-CdR), which is known to promote DNA demethylation in a cell cycle-dependent passive way. In the present work, the process of DNA demethylation following VPA and 5-aza-CdR treatment was studied in HeLa cells. The cells were treated with 1.0 m M VPA for 4 h and then cultivated in the absence of the drug for 24 and 48 h or treated with 5.0 ? M 5-Aza-CdR for 28 h. Immunoassays for 5-methylcytosine (5mC) and 5hmC were performed to evaluate changes in signals related to total DNA methylation and hydroxymethylation. Quantitative PCR assays were used to investigate gene expression levels of TET1 and DNMT1 . Lower fluorescence intensity for methylated DNA in contrast to higher fluorescence intensity for hydroxymethylated DNA was observed in VPAand 5-aza-CdR-treated cells compared to untreated controls, which could be related to an active demethylation induction by both drugs. The DNMT1 mRNA levels were not affected by VPA or 5-aza-CdR treatment. TET1 gene expression decreased after VPA exposure, indicating that other enzymes associated with active demethylation may be involved in this process. Interestingly, 5-azaCdR treatment increased TET1 expression, suggesting that the agents analyzed here may promote active demethylation by different pathways. Taken together, these results help to elucidate the multiple effects performed by these drugs on epigenetic markers.
Valproic acid (VPA) is an important epigenetic drug that works as a histone deacetylase (HDAC) inhibitor, inducing histone hyperacetylation and chromatin unpackaging in several cell types. Currently, DNA demethylation is also being considered part of the VPA action, as shown in HEK293 cells and rat brain cells. This process may occur in a replication-independent active way, possibly with TET proteins participation and 5-hydroxymethylcytosine (5hmC) formation. Recent studies have also detected a 5hmC increase in different cell types after treatment with the well-established DNA methyltransferase (DNMT) inhibitor 5-aza-2′-deoxycytidine (5-aza-CdR), which is known to promote DNA demethylation in a cell cycle-dependent passive way. In the present work, the process of DNA demethylation following VPA and 5-aza-CdR treatment was studied in HeLa cells. The cells were treated with 1.0 m M VPA for 4 h and then cultivated in the absence of the drug for 24 and 48 h or treated with 5.0 μ M 5-Aza-CdR for 28 h. Immunoassays for 5-methylcytosine (5mC) and 5hmC were performed to evaluate changes in signals related to total DNA methylation and hydroxymethylation. Quantitative PCR assays were used to investigate gene expression levels of TET1 and DNMT1 . Lower fluorescence intensity for methylated DNA in contrast to higher fluorescence intensity for hydroxymethylated DNA was observed in VPAand 5-aza-CdR-treated cells compared to untreated controls, which could be related to an active demethylation induction by both drugs. The DNMT1 mRNA levels were not affected by VPA or 5-aza-CdR treatment. TET1 gene expression decreased after VPA exposure, indicating that other enzymes associated with active demethylation may be involved in this process. Interestingly, 5-azaCdR treatment increased TET1 expression, suggesting that the agents analyzed here may promote active demethylation by different pathways. Taken together, these results help to elucidate the multiple effects performed by these drugs on epigenetic markers.
Subject: Células HeLa
Ácido valproico
Epigenética
Metilação de DNA
Cromatina
Reação em cadeia da polimerase
Country: Suíça
Editor: Karger
Citation: Cytogenetic And Genome Research. Karger, v. 148, p. 140 - 140, 2016.
Rights: fechado
Identifier DOI: 10.1159/000446523
Address: https://www.karger.com/Article/Abstract/446523
Date Issue: 2016
Appears in Collections:IB - Artigos e Outros Documentos

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