| Full text | |
| Author(s): |
Rocha, Marina Amorim
;
Breda Veronezi, Giovana Maria
;
Felisbino, Marina Barreto
;
Viccari Gatti, Maria Silvia
;
Tamashiro, Wirla M. S. C.
;
Silveira Mello, Maria Luiza
Total Authors: 6
|
| Document type: | Journal article |
| Source: | SCIENTIFIC REPORTS; v. 9, p. 12-pg., 2019-12-03. |
| Abstract | |
Sodium valproate/valproic acid (VPA), a histone deacetylase inhibitor, and 5-aza-2-deoxycytidine (5-aza-CdR), a DNA methyltransferase 1 (DNMT1) inhibitor, induce DNA demethylation in several cell types. In HeLa cells, although VPA leads to decreased DNA 5-methylcytosine (5mC) levels, the demethylation pathway involved in this effect is not fully understood. We investigated this process using flow cytometry, ELISA, immunocytochemistry, Western blotting and RT-qPCR in G1 phase-arrested and proliferative HeLa cells compared to the presumably passive demethylation promoted by 5-aza-CdR. The results revealed that VPA acts predominantly on active DNA demethylation because it induced TET2 gene and protein overexpression, decreased 5mC abundance, and increased 5-hydroxymethylcytosine (5hmC) abundance, in both G1-arrested and proliferative cells. However, because VPA caused decreased DNMT1 gene expression levels, it may also act on the passive demethylation pathway. 5-aza-CdR attenuated DNMT1 gene expression levels but increased TET2 and 5hmC abundance in replicating cells, although it did not affect the gene expression of TETs at any stage of the cell cycle. Therefore, 5-aza-CdR may also function in the active pathway. Because VPA reduces DNA methylation levels in non-replicating HeLa cells, it could be tested as a candidate for the therapeutic reversal of DNA methylation in cells in which cell division is arrested. (AU) | |
| FAPESP's process: | 14/23842-0 - DNA methylation in HeLa cells after treatment with valproic acid |
| Grantee: | Giovana Maria Breda Veronezi |
| Support Opportunities: | Scholarships in Brazil - Master |
| FAPESP's process: | 15/10356-2 - Action of valproic acid on chromatin structure and function |
| Grantee: | Maria Luiza Silveira Mello |
| Support Opportunities: | Research Projects - Thematic Grants |