title: Involvement of miR-30c and miR-301a in immediate induction of plasminogen activator inhibitor-1 by placental growth factor in human pulmonary endothelial cells.
abstract: PAI-1 (plasminogen activator inhibitor-1) is a key physiological inhibitor of fibrinolysis. Previously, we have reported PlGF (placental growth factor)-mediated transcriptional up-regulation of PAI-1 (SERPINE1) mRNA expression via activation of HIF-1α (hypoxia-inducible factor-1α) and AP-1 (activator protein-1) in HPMVECs (human pulmonary microvascular endothelial cells), which resulted in elevated PAI-1 in humans with SCA (sickle cell anaemia). In the present study, we have identified the role of post-transcriptional mechanism(s) of PlGF-mediated accumulation of PAI-1 mRNA in HPMVECs by examining the role of microRNAs (miRNAs/miRs) in PlGF-induced PAI-1 mRNA stability. Our results show reduced expression of miR-30c and miR-301a, but not of miR-99a, in response to PlGF, which have evolutionarily conserved binding sites in the 3'-UTR (3'-untranslated region) of PAI-1 mRNA. Transfection of anti-miR-30c or anti-miR-301a oligonucleotides resulted in increased PAI-1 mRNA levels, which were increased further with PlGF stimulation. Conversely, overexpression of pre-miR-30c or pre-miR-301a resulted in an attenuation of PlGF-induced PAI-1 mRNA and protein levels. Luciferase reporter assays using wild-type and mutant 3'-UTR constructs confirmed that the PAI-1 3'-UTR is indeed a direct target of miR-30c and miR-301a. Finally, plasma levels of miR-30c and miR-301a were significantly down-regulated in patients with SCA compared with normal controls. These results provide a post-transcriptional regulatory mechanism of PlGF-induced PAI-1 elevation.
authors.affiliation: Department of Biochemistry and Molecular Biology, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA. nitinpat@usc.edu
authors.forename: Nitin
authors.initials: N
authors.lastname: Patel
authors.forename: Stanley M
authors.initials: SM
authors.lastname: Tahara
authors.forename: Punam
authors.initials: P
authors.lastname: Malik
authors.forename: Vijay K
authors.initials: VK
authors.lastname: Kalra
mesh_terms: D020413:3' Untranslated Regions; D000755:Anemia, Sickle Cell / Q000097:blood; D002478:Cells, Cultured; D002648:Child; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000098:blood supply; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000097:blood / Q000502:physiology*; D055806:Microvessels / Q000166:cytology / Q000378:metabolism; D009841:Oligonucleotides / Q000494:pharmacology; D000072483:Placenta Growth Factor; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis* / Q000235:genetics; D011257:Pregnancy Proteins / Q000494:pharmacology / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis
chemical_list: D020413:3' Untranslated Regions; C550735:MIRN301A microRNA, human; C546086:MIRN30b microRNA, human; D035683:MicroRNAs; D009841:Oligonucleotides; C000608036:PGF protein, human; D017395:Plasminogen Activator Inhibitor 1; D011257:Pregnancy Proteins; D012333:RNA, Messenger; D000072483:Placenta Growth Factor