Successful silencing of plasminogen activator inhibitor-1 in human vascular endothelial cells using small interfering RNA.
Thromb Haemost
; 95(5): 857-64, 2006 May.
Article
em En
| MEDLINE
| ID: mdl-16676078
ABSTRACT
Clinical as well as experimental evidence suggests that vascular overexpression of plasminogen activator inhibitor (PAI)-1, the primary physiological inhibitor of both urokinase and tissue-type plasminogen activator, may be involved in the pathophysiology of atherosclerosis and cardiovascular disease. We investigated the feasibility, efficacy and functional effects of PAI-1 gene silencing in human vascular endothelial cells using small interfering RNA. Double-stranded 21 bp-RNA molecules targeted at sequences within the human PAI-1 gene were constructed. Successful siRNA transfection of HUVEC was confirmed using fluorescence microscopy and flow cytometry. One of five candidate siRNA sequences reduced PAI-1 mRNA and protein in a concentration- and time-dependent manner. Suppression of PAI-1 mRNA was detected up to 72 hours after transfection. Moreover, siRNA treatment reduced the activity of PAI-1 released from HUVEC, and prevented the oxLDL- or LPS-induced upregulation of PAI-1 secretion. Importantly, siRNA treatment did not affect the expression of other endothelial-cell markers. Moreover, downregulation of PAI-1 significantly enhanced the ability of endothelial cells to adhere to vitronectin, and this effect could be reversed upon addition of recombinant PAI-1. SiRNA-mediated reduction of PAI-1 expression may be a promising strategy for dissecting the effects of PAI-1 on vascular homeostasis.
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Base de dados:
MEDLINE
Assunto principal:
Endotélio Vascular
/
Inibidor 1 de Ativador de Plasminogênio
/
Inativação Gênica
/
RNA Interferente Pequeno
Limite:
Humans
Idioma:
En
Revista:
Thromb Haemost
Ano de publicação:
2006
Tipo de documento:
Article
País de afiliação:
França