Al. Armesilla et al., Vascular endothelial growth factor activates nuclear factor of activated Tcells in human endothelial cells: a role for tissue factor gene expression, MOL CELL B, 19(3), 1999, pp. 2032-2043
Vascular endothelial growth factor (VEGF) is a potent angiogenic inducer th
at stimulates the expression of tissue factor (TF), the major cellular init
iator of blood coagulation, Here we show that signaling triggered by VEGF i
nduced DNA-binding and transcriptional activities of nuclear factor of acti
vated T cells (NFAT) and AP-1 in human umbilical vein endothelial cells (HU
VECs). VEGF also induced TF mRNA expression and gene promoter activation by
a cyclosporin A (CsA)-sensitive mechanism. As in lymphoid cells, NFAT was
dephosphorylated and translocated to the nucleus upon activation of HUVECs,
and these processes were blocked by CsA. NFAT was involved in the VEGF-med
iated TF promoter activation as evidenced by cotransfection experiments wit
h a dominant negative version of NFAT and site-directed mutagenesis of a ne
wly identified NFAT site within the TF promoter that overlaps with a previo
usly identified kappa B-like site. Strikingly, this site bound exclusively
NFAT not only from nuclear extracts of HUVECs activated by VEGF, a stimulus
that failed to induce NF-kappa B-binding activity, but also from extracts
of cells activated with phorbol eaters and calcium ionophore, a combination
of stimuli that triggered the simultaneous activation of NFAT and NF-kappa
B. These results implicate NFAT in the regulation of endothelial genes by
physiological means and shed light on the mechanisms that switch on the gen
e expression program induced by VEGF and those regulating TF gene expressio
n.