MicroRNAs involved in the regulation of Endothelin-1 gene expression in endothelial cells

2014-08-03We showed that in human dermal microvascular endothelial cell line (HMEC-1), the stability of ET-1 mRNA was increased in response to PlGF treatment. We also observed that in PlGF treated HMEC-1, among several microRNAs (miRNA) which have a potential binding site on ET-1 mRNA and HIF- 1α mRNA, the levels of miR-648, miR-934 and miR-199a-5p were repressed significantly. Furthermore, we showed that among these microRNA candidates, miR-648 attenuated ET-1 mRNA expression, while antimiR-648 increased the expression of ET-1 mRNA under basal and PlGF treated conditions. To further validate this result, we constructed a pGL3-ET-1-3'UTR luciferase reporter. The activity of reporter construct was reduced by miR-648 mimic while antimiR-648 increased the reporter activity, both under basal and PlGF treated conditions. These studies showed miR-648 is involved in the regulation of ET-1 mRNA expression as a result of binding to the 3'UTR region of ET-1 mRNA. This study, for the first time, identified a novel miRNA regulator which functions in the expression of ET-1, and may play an important role in affecting pulmonary hypertension in SCD