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J. Biol. Chem., Vol. 265, Issue 17, 9782-9787, Jun, 1990
DC Crossman, DP Carr, EG Tuddenham, JD Pearson and JH McVey
Tissue factor (TF) is the membrane-bound glycoprotein whose cofactor
activity with factor VIIa causes activation of the extrinsic pathway of
coagulation. The transition of endothelium to a procoagulant state by
agents such as bacterial lipopolysaccharide (LPS) is the result of TF
expression by these cells. The mechanism of TF induction in human umbilical
vein endothelial cells (HUVEC) was investigated in response to LPS and
phorbol 12-myristate 13-O-acetate (PMA). Northern blot analysis of total
RNA from HUVEC showed a rapid rise in TF mRNA levels which was maximal at 2
h and had fallen to low levels by 6 h following both LPS (10 micrograms/ml)
and PMA (10 ng/ml) stimulation. Nuclear-run on experiments showed at most a
2-fold increase in transcription of the TF gene following LPS stimulation
but a 10-fold increase following PMA stimulation. In addition 24-h
pre-incubation with PMA desensitized HUVEC to further PMA exposure, but
caused no alteration in the response to LPS. Cycloheximide (10
micrograms/ml) alone caused induction of TF mRNA. Treatment of cells
previously exposed to LPS for 1 or 4 h with actinomycin D indicated a
12-fold difference in the TF mRNA half-life. Therefore the rapid
accumulation of TF mRNA in HUVEC stimulated by LPS is largely a result of
an increase in mRNA stability rather than an increased rate of
transcription of the gene.
The regulation of tissue factor mRNA in human endothelial cells in response to endotoxin or phorbol ester
Haemostasis Research Group, Clinical Research Centre, Harrow, Middlesex, United Kingdom.
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