MiR-421 and miR-30c Inhibit SERPINE 1 Gene Expression in Human Endothelial Cells

Alexandre Marchand,Pierre-Emmanuel Morange,Anne-Marie Lompré,David-Alexandre Trégouët,Carole Proust,Brian P Chadwick

doi:10.1371/journal.pone.0044532

Abstract

In this work, we assessed whether SERPINE1 expression could be under the influence of microRNAs (miRNAs) predicted to bind the SERPINE1 3′UTR region. We specifically focused on the 3′UTR region harboring a common polymorphism, rs1050955, that have been found associated to SERPINE1 monocyte expression, and investigated whether the presence of different alleles at rs1050955 could modify the miRNAs binding efficiency and affect PAI-1 protein levels. We demonstrated that, in human umbilical vein endothelial cells, both miR-421 and miR-30c directly interacted with PAI-1 mRNA to inhibit the expression of the associated protein. However, these inhibitory mechanisms were independent on the allele present at the rs1050955 locus. We further showed that miR-421 levels correlated with PAI-1 activity in the plasma sample of 40 patients with venous thrombosis. Our results strongly suggest that the regulation of PAI-1 molecule could be under the influence of several miRNAs whose measurement in the plasma of patients could be envisaged as a biomarker for inflammatory and thrombotic disorders.

Highlights

Plasminogen Activator Inhibitor-1 (PAI-1) is a key target in the etiologic and mechanistic study of metabolic syndrome and diseases arising from it
These six miRNAs were tested for co-expression with PAI-1 in a variety of cell lines in which the later is known to be expressed, including human acute monocytic leukemia cells (THP1), human mammary epithelial cells (HMEC), human aortic endothelial cells (HAEC) and human umbilical vein endothelial cells (HUVEC)
By interrogating a public database reporting the results of a genome-wide survey for single nucleotide polymorphisms (SNPs) influencing monocyte gene expressions [10], we observed that the SERPINE1 rs1050955 explained about 4% of the variability of its associated gene

Summary

Introduction

Plasminogen Activator Inhibitor-1 (PAI-1) is a key target in the etiologic and mechanistic study of metabolic syndrome and diseases arising from it. As recently reviewed in Iwaki et al [5], PAI-1 is involved in the physiopathology of nonthrombotic disorders making the disentangling of its underlying regulatory mechanisms a matter of great interest. PAI-1 plasma levels are known to be influenced by both environmental and genetic factors, the latter hypothesized to explain about 30% of their variability [6]. A recent Genome-Wide Association Study (GWAS) conducted in 1,093 individuals from four distinct ethnic groups failed to identify novel robust loci associated with PAI-1 variability [9]. This strongly suggests that a substantial part of the genetic variability remains to be characterized. By interrogating the public GHS_express database [10] reporting the results of a genome wide search for SNPs influencing monocyte gene expressions in a sample of 1,467 healthy individuals, we observed that the rs1050955 mapping the SERPINE1 39UTR region explained ,4% (p,10212) of the SERPINE1 monocyte expression, the rare rs1050955-A allele being associated with decreased SERPINE1 expression

Methods

Results

Conclusion