Identification of target proteins of shear stress‐related miRNAs in endothelial cells

Abstract

Atherosclerosis is an inflammatory disease that occurs preferentially at particular areas of disturbed flow characterized by low and oscillatory wall shear stress (OS) in branched or curved arteries. In contrast, straight arterial regions are exposed to high and stable laminar shear stress(LS) and are well protected from atherosclerosis.To understand the molecular mechanisms by which shear stress elicits proatherogenic phenotype, we have performed miRNA microarray using in vitro cone and plate model and in vivo mouse partial carotid artery ligation model. As a result, we identified miR663 and miR712 wasas the most induced miRNAs in Human umbilical vein endothelial cells(HUVECs) and mouse aortic endothelial cells(MAECs).Furthermore, we employed 2D‐DIGE in order to identify the target genes that are regulated by shear sensitive miR663 and miR712. As a result, in HUVECs, we were able to find 4 spots from soluble fraction(S‐100), and 3 spots from heavy membrane(HM) fraction, and 2 spots from light membrane(LM) fraction, which were downregulated upon pre‐miR 663 overexpression by minimum fold of 1.2. And in MAECs we found 4 spots from HM fraction which were upregulated upon anti miR712 LNA by minimum fold of 1.2. The proteins were analyzed further by mass spectrometry.Therefore the proteomic technology allowed as to identify novel targets of miRNA in endothelial cells.