MicroRNA-223 inhibits tissue factor expression in vascular endothelial cells

Atherosclerosis. 2014 Dec;237(2):514-20. doi: 10.1016/j.atherosclerosis.2014.09.033. Epub 2014 Oct 18.


Objective: Atherosclerosis is a chronic inflammatory process, in which vascular endothelial cells (ECs) become dysfunctional owing to the effects of chemical substances, such as inflammatory factor and growth factors. Tissue factor (TF) expression is induced by the above chemical substances in activated ECs. TF initiates thrombosis on disrupted atherosclerotic plaques which plays an essential role during the onset of acute coronary syndromes (ACS). Increasing evidences suggest the important role of microRNAs as epigenetic regulators of atherosclerotic disease. The aim of our study is to identify if microRNA-223 (miR-223) targets TF in ECs.

Methods and results: Bioinformatic analysis showed that TF is a target candidate of miR-223. Western blotting analysis revealed that tumor necrosis factor α (TNF-α) increased TF expression in aorta of C57BL/6J mice and cultured ECs (EA.hy926 cells and HUVEC) after 4 h treatment. In TNF-α treated ECs, TF mRNA was also increased measured by real-time PCR. Real-time PCR results showed that miR-223 levels were downregulated in TNF-α-treated aorta of C57BL/6J mice and cultured ECs. Transfection of ECs with miR-223 mimic or miR-223 inhibitor modified TF expression both in mRNA and protein levels. Luciferase assays confirmed that miR-223 suppressed TF expression by binding to the sequence of TF 3'-untranslated regions (3'UTR). TF procoagulant activity was inhibited by overexpressing miR-223 with or without TNF-α stimulation.

Conclusions: MiR-223-mediated suppression of TF expression provides a novel molecular mechanism for the regulation of coagulation cascade, and suggests a clue against thrombogenesis during the process of atherosclerotic plaque rupture.

Keywords: Atherosclerosis; TNF-α; Thrombogenesis; Tissue factor; microRNA-223.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aorta / metabolism
  • Aorta / pathology
  • Base Sequence
  • Binding Sites
  • Blood Coagulation
  • Cells, Cultured
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Gene Expression Regulation*
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Inflammation
  • Male
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics
  • MicroRNAs / physiology*
  • Molecular Sequence Data
  • Plaque, Atherosclerotic / metabolism
  • Thromboplastin / antagonists & inhibitors
  • Thromboplastin / metabolism*
  • Tumor Necrosis Factor-alpha / metabolism


  • MIRN223 microRNA, human
  • MIRN223 microRNA, mouse
  • MicroRNAs
  • Tumor Necrosis Factor-alpha
  • Thromboplastin