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J Biol Chem, Vol. 274, Issue 46, 32795-32802, November 12, 1999
From the Departments of Cell Biology and Cardiology, Cleveland
Clinic Foundation, Cleveland, Ohio 44195
Tissue factor, in association with factor VIIa,
initiates the coagulation cascade. We studied the influences of two
pathophysiological stimuli, native (unmodified) and oxidized low
density lipoprotein, on tissue factor gene expression in a cell
important in vascular remodeling and vascular diseases, the smooth
muscle cell. Our results demonstrated that both lipoproteins
significantly induced tissue factor gene expression in rat aortic
smooth muscle cells; oxidized low density lipoprotein was slightly more
potent. Both lipoproteins increased tissue factor mRNA in a
concentration- and time-dependent manner. Results from
nuclear run-on assays and mRNA stability experiments indicated that
increased tissue factor mRNA accumulation in response to the
lipoproteins was principally controlled at the transcriptional level.
By using lipid extracts of low density lipoprotein or methylation of
the intact lipoprotein to block receptor recognition, we showed that
this lipoprotein induced tissue factor mRNA via both
receptor-independent and receptor-augmented pathways. Transfection
studies using a series of deleted tissue factor promoters revealed that
a 
143- to +106-base pair region of the rat tissue factor promoter
contained regulatory elements required for lipoprotein-mediated
induction. Electrophoretic mobility shift assays showed that the
binding activities of the transcription factor Egr-1, but not Sp1, were
markedly elevated in response to these lipoproteins. Transfection of
site-directed mutants of the tissue factor (TF) promoter demonstrated
that not only Egr-1 but also Sp1 cis-acting elements in the
TF (143) promoter construct were necessary for optimal TF gene
induction. Our data show for the first time that both low density
lipoprotein and oxidized low density lipoprotein induce tissue factor
gene expression in smooth muscle cells and that this tissue factor gene
expression is mediated by both Egr-1 and Sp1 transcription factors.
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