Poomipark, N,, Flatley, J,E., Hill, M.H. et al. (4 more authors) (2016) Methyl donor status influences DNMT expression and global DNA methylation in cervical cancer cells. Asia Pacific Journal of Cancer Prevention, 17 (7). pp. 3213-3222. ISSN 1513-7368
Abstract
Background: Methyl donor status influences DNA stability and DNA methylation although little is known about effects on DNA methyltransferases. The aim of this study was to determine whether methyldonor status influences DNA methyltransferase (Dnmt) gene expression in cervical cancer cells, and if so, whether there are associated effects on global DNA methylation.
Materials and Methods: The human cervical cancer cell line, C4 II, was grown in complete medium and medium depleted of folate (FM) and folate and methionine (FM). Growth rate, intracellular folate, intracellular methionine and homocysteine in the extracellular medium were measured to validate the cancer cell model of methyl donor depletion. Dnmt expression was measured by qRT PCR using relative quantification and global DNA methylation was measured using a flow cytometric method.
Results: Intracellular folate and methionine concentrations were significantly reduced after growth in depleted media. Growth rate was also reduced in response to methyl donor depletion. Extracellular homocysteine was raised compared with controls, indicating disturbance to the methyl cycle. Combined folate and methionine depletion led to a significant downregulation of Dnmt3a and Dnmt3b; this was associated with an 18% reduction in global DNA methylation compared with controls. Effects of folate and methionine depletion on Dnmt3a and 3b expression were reversed by transferring depleted cells to complete medium.
Conclusions: Methyl donor status can evidently influence expression of Dnmts in cervical cancer cells, which is associated with DNA global hypomethylation. Effects on Dnmt expression are reversible, suggesting reversible modulating effects of dietary methyl donor intake on gene expression, which may be relevant for cancer progression.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2016 APOCP. |
Keywords: | Cervical cancer; folate; methionine; DNA methylation; DNA methyltransferase |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > School of Health and Related Research (Sheffield) > ScHARR - Sheffield Centre for Health and Related Research The University of Sheffield > Sheffield Teaching Hospitals |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 05 Dec 2019 11:39 |
Last Modified: | 05 Dec 2019 11:39 |
Published Version: | http://journal.waocp.org/?sid=Entrez:PubMed&id=pmi... |
Status: | Published |
Publisher: | Asian Pacific Organization for Cancer Prevention (APOCP) |
Refereed: | Yes |
Identification Number: | 10.14456/apjcp.2016.78/APJCP.2016.17.7.3213 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:154207 |