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Volume 270,
Number 41,
Issue of October 13, 1995 pp. 24188-24196
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Fibroblast Growth
Factor-2 Can Mediate Cell Attachment by Linking Receptors and Heparan
Sulfate Proteoglycans on Neighboring Cells
(Received for publication, April 27, 1995; and in revised form, July 18, 1995)
Christian
Richard,
John
P.
Liuzzo,
David
Moscatelli
The myeloid 32D cell line, which grows in suspension and does
not express FGF receptors or heparan sulfate proteoglycans, was
transfected with the cDNA encoding FGF receptor-1 (32D-flg cells). When
co-cultured with glutaraldehyde-fixed Chinese hamster ovary (CHO)
cells, the 32D-flg cells remained in suspension in the absence of FGF-2
but attached to the CHO monolayer in the presence of 10 ng/ml FGF-2. In
contrast, 32D cells transfected with the vector alone did not attach to
the CHO monolayer in the presence of FGF-2. FGF-2-dependent attachment
of 32D-flg cells was prevented by inclusion of 10 µg/ml heparin in
the incubation medium and was diminished when CHO mutants in
glycosaminoglycan synthesis or wild-type CHO cells treated with
heparinase were used, indicating that the attachment occurred through
FGF-2 interactions with heparan sulfates on the CHO cells. Attachment
of 32D-flg cells to wild-type CHO cells was half-maximal at 0.4 ng/ml
FGF-2 and was also observed with FGF-1 but not FGF-4. 32D-flg cells
also attached to living CHO cells in a FGF-2-dependent manner, but
attachment was transient at 37 °C. Induction of new proteins was
not required for FGF-2-dependent attachment, since attachment occurred
when the co-cultures were incubated at 4 °C and when the 32D-flg
cells were preincubated with cycloheximide. FGF-2-dependent attachment
of 32D-flg cells was also observed with Balb/C 3T3, NIH 3T3, and bovine
capillary endothelial cells. We conclude that attachment is due to
FGF-2 binding simultaneously to receptors on the 32D-flg cells and
heparan sulfates on the CHO monolayers; thus, the FGF-2 acts as a
bridge between receptorexpressing cells and heparan sulfate-bearing
cells. In addition, induction of DNA synthesis in 32D-flg cells in
response to FGF-2 was potentiated by the CHO-associated heparan
sulfates to the same extent as by soluble heparin, indicating that this
interaction has functional significance.

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Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.
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