MiRNA-155 targets myosin light chain kinase and modulates actin cytoskeleton organization in endothelial cells

Am J Physiol Heart Circ Physiol. 2014 Apr 15;306(8):H1192-203. doi: 10.1152/ajpheart.00521.2013. Epub 2014 Jan 31.

Abstract

Previously, we identified a microRNA (miRNA) signature for endothelial cells (ECs) subjected to unidirectional shear stress (USS). MiR-155, a multifunctional miRNA that has been implicated in atherosclerosis, was among the shear stress-responsive miRNAs. Here, we examined the role of miR-155 in modulating EC phenotype and function. In vitro, increased miR-155 levels in human ECs induced changes in morphology and filamentous (F)-actin organization. In addition, ECs transfected with miR-155 mimic were less migratory and less proliferative and had less apoptosis compared with control ECs. In mouse aorta, miR-155 expression was increased in the intima of thoracic aorta, where blood flow produces steady and unidirectional shear stress, compared with the intima of the lower curvature of the aortic arch, which is associated with oscillatory and low shear stress. These differences in miR-155 expression were associated with distinct changes in EC morphology and F-actin. The effects of miR-155 in vitro were mediated through suppression of two key regulators of the EC cytoskeleton organization: RhoA and myosin light chain kinase (MYLK). A novel direct interaction between miR-155 and the MYLK 3'UTR was verified by luciferase-MYLK 3'UTR reporter assays. Furthermore, the intensity of immunofluorescence staining for RhoA and MYLK in mouse aorta correlated inversely with miR-155 expression. In conclusion, a prominent effect of the multifunctional miR-155 in ECs is modulation of phenotype through alterations in RhoA, MYLK expression, and actin cytoskeleton organization.

Keywords: cytoskeleton; endothelial cell; microRNA; shear stress; stress fibers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actin Cytoskeleton / ultrastructure*
  • Animals
  • Aorta / chemistry
  • Aorta / metabolism
  • Atherosclerosis
  • Endothelial Cells / enzymology*
  • Endothelial Cells / ultrastructure*
  • Fluorescent Antibody Technique
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics
  • MicroRNAs / physiology*
  • Myosin-Light-Chain Kinase / analysis
  • Myosin-Light-Chain Kinase / metabolism*
  • Phenotype
  • Transfection
  • rhoA GTP-Binding Protein / analysis

Substances

  • MIRN155 microRNA, human
  • MicroRNAs
  • Myosin-Light-Chain Kinase
  • rhoA GTP-Binding Protein