The microRNA miR-134-5p induces calcium deposition by inhibiting histone deacetylase 5 in vascular smooth muscle cells

J Cell Mol Med. 2020 Sep;24(18):10542-10550. doi: 10.1111/jcmm.15670. Epub 2020 Aug 11.

Abstract

Calcium deposition in vascular smooth muscle cells (VSMCs) is a form of ectopic ossification in blood vessels. It can result in rigidity of the vasculature and an increase in cardiac events. Here, we report that the microRNA miR-134-5p potentiates inorganic phosphate (Pi)-induced calcium deposition in VSMCs by inhibiting histone deacetylase 5 (HDAC5). Using miRNA microarray analysis of Pi-treated rat VSMCs, we first selected miR-134-5p for further evaluation. Quantitative RT-PCR confirmed that miR-134-5p was increased in Pi-treated A10 cells, a rat VSMC line. Transfection of miR-134-5p mimic potentiated the Pi-induced increase in calcium contents. miR-134-5p increased the amounts of bone runt-related transcription factor 2 (RUNX2) protein and bone morphogenic protein 2 (BMP2) mRNA in the presence of Pi but decreased the expression of osteoprotegerin (OPG). Bioinformatic analysis showed that the HDAC5 3'untranslated region (3'UTR) was one of the targets of miR-134-5p. The luciferase construct containing the 3'UTR of HDAC5 was down-regulated by miR-134-5p mimic in a dose-dependent manner in VSMCs. Overexpression of HDAC5 mitigated the calcium deposition induced by miR-134-5p. Our results suggest that a Pi-induced increase of miR-134-5p may cause vascular calcification through repression of HDAC5.

Keywords: miR-134-5p; histone deacetylase 5; microRNA; vascular calcification; vascular smooth muscle cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Aorta, Thoracic / cytology
  • Calcium / metabolism*
  • Cell Line
  • Computer Simulation
  • Core Binding Factor Alpha 1 Subunit / biosynthesis
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / immunology
  • Down-Regulation
  • Gene Expression Regulation
  • Genes, Reporter
  • Histone Deacetylases / biosynthesis
  • Histone Deacetylases / drug effects*
  • Histone Deacetylases / genetics
  • MicroRNAs / genetics
  • MicroRNAs / physiology*
  • Microarray Analysis
  • Muscle, Smooth, Vascular / cytology
  • Myocytes, Smooth Muscle / metabolism*
  • Osteoprotegerin / biosynthesis
  • Osteoprotegerin / genetics
  • Phosphates / toxicity
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Fusion Proteins / metabolism
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Transfection
  • Vascular Calcification / etiology*
  • Vascular Calcification / genetics
  • Vascular Calcification / metabolism
  • Vascular Calcification / prevention & control

Substances

  • 3' Untranslated Regions
  • Core Binding Factor Alpha 1 Subunit
  • MIRN134 microRNA, rat
  • MicroRNAs
  • Osteoprotegerin
  • Phosphates
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Runx2 protein, rat
  • Tnfrsf11b protein, rat
  • Hdac5 protein, rat
  • Histone Deacetylases
  • Calcium