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Dynamic Enhancer Methylation - A Previously Unrecognized Switch for Tissue-Type Plasminogen Activator Expression.

Artikel i vetenskaplig tidskrift
Författare Mia Magnusson
Emma Xuchun Lu
Pia Larsson
Erik Ulfhammer
Niklas Bergh
Helena Carén
Sverker Jern
Publicerad i PloS one
Volym 10
Nummer/häfte 10
Sidor e0141805
ISSN 1932-6203
Publiceringsår 2015
Publicerad vid Wallenberglaboratoriet
Sahlgrenska Cancer Center
Institutionen för biomedicin, avdelningen för patologi
Institutionen för medicin, avdelningen för molekylär och klinisk medicin
Sidor e0141805
Språk en
Länkar dx.doi.org/10.1371/journal.pone.014...
Ämnesord louise.gracanin@vgregion.se
Ämneskategorier Kardiovaskulär medicin

Sammanfattning

Tissue-type plasminogen activator (t-PA), which is synthesized in the endothelial cells lining the blood vessel walls, is a key player in the fibrinolytic system protecting the circulation against occluding thrombus formation. Although classical gene regulation has been quite extensively studied in order to understand the mechanisms behind t-PA regulation, epigenetics, including DNA methylation, still is a largely unexplored field. The aim of this study was to establish the methylation pattern in the t-PA promoter and enhancer in non-cultured compared to cultured human umbilical vein endothelial cells (HUVECs), and to simultaneously examine the level of t-PA gene expression. Bisulphite sequencing was used to evaluate the methylation status, and real-time RT-PCR to determine the gene expression level. While the t-PA promoter was stably unmethylated, we surprisingly observed a rapid reduction in the amount of methylation in the enhancer during cell culturing. This demethylation was in strong negative correlation with a pronounced (by a factor of approximately 25) increase in t-PA gene expression levels. In this study, we show that the methylation level in the t-PA enhancer appears to act as a previously unrecognized switch controlling t-PA expression. Our findings, which suggest that DNA methylation is quite dynamic, have implications also for the interpretation of cell culture experiments in general, as well as in a wider biological context.

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http://www.gu.se/forskning/publikation/?publicationId=225480
Utskriftsdatum: 2019-07-18