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Molecular and Cellular Biochemistry

, Volume 412, Issue 1–2, pp 111–117 | Cite as

Thrombospondin-1 differentially regulates microRNAs in vascular smooth muscle cells

  • Kristopher G. MaierEmail author
  • Brian Ruhle
  • Jeffrey J. Stein
  • Karen L. Gentile
  • Frank A. Middleton
  • Vivian Gahtan
Article

Abstract

Thrombospondin-1 (TSP-1) is an important regulator of vascular smooth muscle cell (VSMC) physiology and gene expression. MicroRNAs (microRNA), small molecules that regulate protein translation, have emerged as potent regulators of cell function. MicroRNAs have been shown to be involved in intimal hyperplasia, atherosclerosis, and upregulated in the vasculature in diabetes. The purpose of this study was to identify microRNAs regulated by TSP-1 in vascular smooth muscle cells (VSMCs). Human VSMCs were treated for 6 h with basal media or TSP-1 both supplemented with 0.2 % FBS. Cells were then snap frozen and RNA extracted. An Affymetrix GeneChip microRNA array analysis was performed in triplicate on three separate collections. Confirmatory qrtPCR was performed. Data were analyzed by ANOVA or t test, with significance set at p < 0.05. MicroRNAs identified were subjected to KEGG pathway analysis using the DIANA tools miRPath online tool. TSP-1 upregulated 22 microRNAs and downregulated 18 microRNAs in VSMCs (p < 0.05). The most upregulated microRNA was miR-512-3p (45.12 fold). The microRNA most downregulated by TSP-1 was miR-25-5p, which was decreased by 9.61. Of note, five members of the mir-17–92 cluster were downregulated. KEGG analysis revealed that thirty-three cellular signaling pathways were impacted by these microRNAs and that nine pathways were relevant to vascular disease. MicroRNAs regulate protein expression at the level of translation and may represent a significant mechanism by which TSP-1 regulates VSMC function. Several of the microRNAs identified have a role in vascular function. The miR-17–92 cluster family, which was found to exhibit reduced expression in this study, is known to be involved in angiogenesis and vascular function. TSP-1 regulates multiple microRNAs in VSMCs adding a new layer of complexity to TSP-1 regulation of VSMC function.

Keywords

MicroRNA Thrombospondins Gene expression profiling Vascular smooth muscle 

Notes

Funding

This research was supported by a Grant from the Central New York Research Fund and a VA Merit award #BX001243.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest with the materials in this manuscript.

Supplementary material

11010_2015_2614_MOESM1_ESM.xlsx (12 kb)
Supplementary material 1 (XLSX 12 kb)

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Copyright information

© Springer Science+Business Media New York (outside the USA) 2016

Authors and Affiliations

  • Kristopher G. Maier
    • 1
    • 3
    Email author
  • Brian Ruhle
    • 1
    • 3
  • Jeffrey J. Stein
    • 1
    • 3
  • Karen L. Gentile
    • 2
  • Frank A. Middleton
    • 2
  • Vivian Gahtan
    • 1
    • 3
  1. 1.Division of Vascular Surgery and Endovascular ServicesSUNY Upstate Medical University College of MedicineSyracuseUSA
  2. 2.Department of Neuroscience and PhysiologySUNY Upstate Medical UniversitySyracuseUSA
  3. 3.Department of Veterans Affairs VA Healthcare Network Upstate New York at SyracuseSyracuseUSA

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