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    Mapping the methylation status of the miR-145 promoter in saphenous vein smooth muscle cells from individuals with type 2 diabetes

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    Publication date
    2016
    Author
    Riches-Suman, Kirsten
    Huntriss, J.
    Keeble, C.
    Wood, I.C.
    O'Regan, D.J.
    Turner, N.A.
    Porter, K.E.
    Keyword
    miR-145; DNA methylation; Type 2 diabetes; Smooth muscle cells; Pyrosequencing; Saphenous vein
    Rights
    © The Authors 2016. Distributed under the terms of the Creative Commons Attribution 3.0 License (http://www.creativecommons.org/licenses/by/3.0/) which permits any use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
    Peer-Reviewed
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    Abstract
    Type 2 diabetes mellitus prevalence is growing globally, and the leading cause of mortality in these patients is cardiovascular disease. Epigenetic mechanisms such as microRNAs (miRs) and DNA methylation may contribute to complications of type 2 diabetes mellitus. We discovered an aberrant type 2 diabetes mellitus–smooth muscle cell phenotype driven by persistent up-regulation of miR-145. This study aimed to determine whether elevated expression was due to changes in methylation at the miR-145 promoter. Smooth muscle cells were cultured from saphenous veins of 22 non-diabetic and 22 type 2 diabetes mellitus donors. DNA was extracted, bisulphite treated and pyrosequencing used to interrogate methylation at 11 CpG sites within the miR-145 promoter. Inter-patient variation was high irrespective of type 2 diabetes mellitus. Differential methylation trends were apparent between non-diabetic and type 2 diabetes mellitus–smooth muscle cells at most sites but were not statistically significant. Methylation at CpGs −112 and −106 was consistently lower than all other sites explored in non-diabetic and type 2 diabetes mellitus–smooth muscle cells. Finally, miR-145 expression per se was not correlated with methylation levels observed at any site. The persistent up-regulation of miR- 145 observed in type 2 diabetes mellitus–smooth muscle cells is not related to methylation at the miR-145 promoter. Crucially, miR-145 methylation is highly variable between patients, serving as a cautionary note for future studies of this region in primary human cell types.
    URI
    http://hdl.handle.net/10454/11032
    Version
    Published version
    Citation
    Riches-Suman K, Huntriss J, Keeble C et al (2016) Mapping the methylation status of the miR-145 promoter in saphenous vein smooth muscle cells from individuals with type 2 diabetes. Diabetes & Vascular Disease Research. 14 (2): 122-129.
    Link to publisher’s version
    http://dx.doi.org/10.1177/1479164116677968
    Type
    Article
    Collections
    Life Sciences Publications

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