TY - JOUR
T1 - Serum stimulation of quiescent human fibroblasts induces the synthesis of tissue factor mRNA followed by the appearance of tissue factor antigen and procoagulant activity
AU - Bloem, Laura J.
AU - Chen, Lisa
AU - Konigsberg, William H.
AU - Bach, Ronald R
PY - 1989/5
Y1 - 1989/5
N2 - Expression of the gene for tissue factor, the cell‐surface initiator of blood coagulation, was examined by stimulating growth‐arrested human fibroblasts with serum and measuring changes in the cellular content of tissue factor mRNA, antigen, and activity. Maximum tissue factor mRNA levels were reached within 1 h following serum induction and slowly declined to basal levels from 24 to 48 h after stimulation. The appearance of the tissue factor mRNA was followed by an increase in tissue factor antigen and activity. The parallel rise in antigenically positive protein and procoagulant activity was first observed about 2 h after serum stimulation with a peak at 12 h followed by a slow decline during the next 36 h. The serum‐induced synthesis of the tissue factor mRNA was independent of de novo protein synthesis as demonstrated by the increased tissue factor mRNA levels generated in the presence of cycloheximide. The results of this study suggest that the synthesis of tissue factor in human fibroblasts i s regulated principally at the level of transcription. In one strain of fibroblasts the activity/antigen ratio, during the period of maximum synthesis, was indistinguishable from that of tissue factor which had been immunoaffinity purified from human brain and reconstituted into phospholipid vesicles. However, during serum starvation the activity/antigen ratio in these cells was significantly reduced. Western blot analysis revealed that in serum‐starved cells there was an accumulation of truncated forms of the tissue factor antigen while in the serum‐stimulated cells only the full‐length antigen was observed.
AB - Expression of the gene for tissue factor, the cell‐surface initiator of blood coagulation, was examined by stimulating growth‐arrested human fibroblasts with serum and measuring changes in the cellular content of tissue factor mRNA, antigen, and activity. Maximum tissue factor mRNA levels were reached within 1 h following serum induction and slowly declined to basal levels from 24 to 48 h after stimulation. The appearance of the tissue factor mRNA was followed by an increase in tissue factor antigen and activity. The parallel rise in antigenically positive protein and procoagulant activity was first observed about 2 h after serum stimulation with a peak at 12 h followed by a slow decline during the next 36 h. The serum‐induced synthesis of the tissue factor mRNA was independent of de novo protein synthesis as demonstrated by the increased tissue factor mRNA levels generated in the presence of cycloheximide. The results of this study suggest that the synthesis of tissue factor in human fibroblasts i s regulated principally at the level of transcription. In one strain of fibroblasts the activity/antigen ratio, during the period of maximum synthesis, was indistinguishable from that of tissue factor which had been immunoaffinity purified from human brain and reconstituted into phospholipid vesicles. However, during serum starvation the activity/antigen ratio in these cells was significantly reduced. Western blot analysis revealed that in serum‐starved cells there was an accumulation of truncated forms of the tissue factor antigen while in the serum‐stimulated cells only the full‐length antigen was observed.
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U2 - 10.1002/jcp.1041390226
DO - 10.1002/jcp.1041390226
M3 - Article
C2 - 2715194
AN - SCOPUS:0024507992
SN - 0021-9541
VL - 139
SP - 418
EP - 423
JO - Journal of cellular physiology
JF - Journal of cellular physiology
IS - 2
ER -