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Blood, Vol. 95 No. 11 (June 1), 2000:
pp. 3387-3395
Up-regulation of vascular endothelial growth factor receptor Flt-1
after endothelial denudation: role of transcription factor Egr-1
Felipe Vidal,
Julián Aragonés,
Arántzazu Alfranca, and
Manuel O. de
Landázuri
From the Servicio de Inmunología, Hospital de la Princesa,
Universidad Autónoma de Madrid, Madrid, Spain.
Vascular endothelial growth factor (VEGF) is highly expressed in
vascular remodeling processes and accelerates reendothelialization after mechanical denudation. Two VEGF tyrosine kinase receptors have
been reported fms-like-tyrosine kinase-1 (Flt-1) and kinase domain region (KDR). Little is known about the regulation of the expression of these receptors after vascular injury. Herein, we have
analyzed the expression of Flt-1 after mechanical denudation of primary
cultures of endothelial cells, which has been considered a useful in
vitro model to study endothelium responses to vascular injury. After
denudation, the Flt-1 protein and mRNA levels are clearly up-regulated,
and transient transfection experiments showed a strong induction of the
flt-1 promoter-dependent transcription. Analysis
of the flt-1 promoter sequence revealed the presence of a
putative binding site for the early growth response factor-1 (Egr-1) at
positions  24 to 16. Electrophoretic mobility shift and supershift assays showed that Egr-1 was able to bind to this DNA
sequence, and cotransfection of the flt-1 promoter reporter plasmid with an Egr-1 expression vector resulted in enhancement of its
transcriptional activity. Furthermore, the mutation of the Egr-1
binding site markedly reduced the denudation-induced flt-1
promoter activity. These data demonstrate that Flt-1 is up-regulated
after endothelial denudation and that Egr-1 plays a relevant role in
this process.
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