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Rapid and specific hypomethylation of enhancers in endothelial cells during adaptation to cell culturing.

Artikel i vetenskaplig tidskrift
Författare Mia Magnusson
Pia Larsson
Emma Xuchun Lu
Niklas Bergh
Helena Carén
Sverker Jern
Publicerad i Epigenetics
Volym 11
Nummer/häfte 8
Sidor 614-624
ISSN 1559-2308
Publiceringsår 2016
Publicerad vid Wallenberglaboratoriet
Sahlgrenska Cancer Center
Institutionen för biomedicin, avdelningen för patologi
Institutionen för medicin, avdelningen för molekylär och klinisk medicin
Sidor 614-624
Språk en
Länkar dx.doi.org/10.1080/15592294.2016.11...
Ämneskategorier Kardiovaskulär medicin

Sammanfattning

Epigenetics, including DNA methylation, is one way for a cell to respond to the surrounding environment. Traditionally, DNA methylation has been perceived as a quite stable modification; however, lately, there have been reports of a more dynamic CpG methylation that can be affected by, for example, long-term culturing. We recently reported that methylation in the enhancer of the gene encoding the key fibrinolytic enzyme tissue-type plasminogen activator (t-PA) was rapidly erased during cell culturing. In the present study we used sub-culturing of human umbilical vein endothelial cells (HUVECs) as a model of environmental challenge to examine how fast genome-wide methylation changes can arise. To assess genome-wide DNA methylation, the Infinium HumanMethylation450 BeadChip was used on primary, passage 0, and passage 4 HUVECs. Almost 2% of the analyzed sites changed methylation status to passage 4, predominantly displaying hypomethylation. Sites annotated as enhancers were overrepresented among the differentially methylated sites (DMSs). We further showed that half of the corresponding genes concomitantly altered their expression, most of them increasing in expression. Interestingly, the stroke-related gene HDAC9 increased its expression several hundredfold. This study reveals a rapid hypomethylation of CpG sites in enhancer elements during the early stages of cell culturing. As many methods for methylation analysis are biased toward CpG rich promoter regions, we suggest that such methods may not always be appropriate for the study of methylation dynamics. In addition, we found that significant changes in expression arose in genes with enhancer DMSs. HDAC9 displayed the most prominent increase in expression, indicating, for the first time, that dynamic enhancer methylation may be central in regulating this important stroke-associated gene.

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