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J Biol Chem, Vol. 274, Issue 7, 4400-4411, February 12, 1999
From the Re-expression of EGR-1 in fibrosarcoma HT1080
suppresses transformation including tumorigenicity (Huang, R.-P., Liu,
C., Fan, Y., Mercola, D., and Adamson, E. (1995) Cancer
Res. 55, 5054-5062) owing in part to up-regulation of the
transforming growth factor (TGF)-
The Transcription Factor EGR-1 Suppresses Transformation of Human
Fibrosarcoma HT1080 Cells by Coordinated Induction of Transforming
Growth Factor-
1, Fibronectin, and Plasminogen Activator
Inhibitor-1
,
,**
Sidney Kimmel Cancer Center,
San Diego, California 92121, the § Department of
Immunology, The Scripps Research Institute,
La Jolla, California 92037, the ¶ Burnham Institute,
La Jolla, California 92037, the Northwest Cancer Research
Center, Seattle, Washington 98125, and the ** Center for Molecular
Genetics, University of California at San Diego,
La Jolla, California 92093
1 promoter by EGR-1 which
suppresses growth by an autocrine mechanism (Liu, C., Adamson, E., and
Mercola, D. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 11831-11836). Here we show that enhanced cell attachment contributes
to the suppression via increased secretion of fibronectin (FN) and also
of plasminogen activator inhibitor-1 (PAI-1). The secretion of FN and
PAI-1 is strongly correlated with EGR-1 expression
(RPEARSON = 0.971 and 0.985, respectively).
Addition of authentic TGF-1 to parental cells greatly stimulated
secretion of PAI-1 but not FN, whereas addition of TGF- antibody or
lipofection with specific antisense TGF-1 oligonucleotides to EGR-1-regulated cells completely inhibits the
secretion of PAI-1 but not FN. However, in gel mobility shift assays
pure EGR-1 or nuclear extracts of EGR-1-regulated cells specifically
bind to two GC-rich elements of the human FN promoter at
positions 
75/52 and 4/+18, indicating that the increased secretion of FN is likely due to direct up-regulation by EGR-1. Moreover, adhesion was greatly enhanced in EGR-1-regulated cells and
was reversed by treatment with Arg-Gly-Asp (RGD) or PAI-1 antibody
indicating that the secreted proteins are functional. We conclude that
EGR-1 regulates the coordinated expression of gene products important
for cell attachment ("oikis" factor) and normal growth control.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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