Growth factors stimulate sustained cell migration as well as inducing select acute motility-related events such as membrane ruffling and disruption of focal adhesions. However, an in-depth understanding of the characteristics of sustained migration that are regulated by growth factor signals is lacking: how the biochemical signals are related to physical processes underlying locomotion, and how these events are coordinately influenced by interplay between growth factor and matrix substratum signals. To address these issues, we studied sustained migration of NR6 fibroblasts on a complex human matrix substratum, Amgel, comparing effects of epidermal growth factor (EGF) treatment across a range of Amgel levels. In the absence of EGF, cell migration speed and directional persistence are relatively independent of Amgel level, whereas in the presence of EGF speed is increased at intermediate Amgel levels but not at low and high Amgel levels while directional persistence is decreased at intermediate but not at low and high Amgel levels. The net effect of EGF is to increase the frequency of changes in the cell direction, and at the same time to slightly increase the path-length and thereby greatly enhance random dispersion of cells. Despite increasing migration speed during long-term sustained migration EGF treatment does not lead to significantly increased absolute rates of membrane extension in contrast to its well-known elicitation of membrane ruffling in the short term. However, EGF treatment does decrease cell spread area, yielding an apparent enhancement of specific membrane extension rate, i.e. normalized to cell spread area. Cell movement speed and directional persistence are thus, respectively, directly related and indirectly related to the increase in specific membrane extension rate (alternatively, the decrease in cell spread area) induced by EGF treatment during sustained migration. These results indicate that growth factor and matrix substrata coordinately regulate sustained cell migration through combined governance of underlying physical processes.

REFERENCES

Aaronson
S. A.
(
1991
).
Growth factors and cancer.
Science
254
,
1146
1153
Ando
Y.
,
Jensen
P. J.
(
1993
).
Epidermal growth factor and insulin-like growth factor I enhance keratinocyte migration.
J. Invest. Dermatol
100
,
633
639
Blay
J.
,
Brown
K. D.
(
1985
).
Epidermal growth factor promotes the chemotactic migration of cultured rat intestinal epithelial cells.
J. Cell. Physiol
124
,
107
112
Bornfeldt
K. E.
,
Raines
E. W.
,
Nakano
T.
,
Graves
L. M.
,
Krebs
E. G.
,
Ross
R.
(
1994
).
Insulin-like growth factor-1 and platelet-derived growth factor-BB induce directed migration of human arterial smooth muscle cells via signalling pathways that are distinct from those of proliferation.
J. Clin. Invest
93
,
1266
1274
Bornfeldt
K. E.
,
Graves
L. M.
,
Raines
E. W.
,
Igarashi
Y.
,
Wayman
G.
,
Yamamura
S.
,
Yatomi
Y.
,
Sidhu
J. S.
,
Krebs
E. G.
,
Hakomori
S.-I.
, et al. 
(
1995
).
Sphingosine-1-phosphate inhibits PDGF-induced chemotaxis of human arterial smooth muscle cells: spatial and temporal modulation of PDGF chemotactic signal transduction.
J. Cell Biol
130
,
193
206
Cha
D.
,
O'Brien
P.
,
O'Toole
E. A.
,
Woodley
D. T.
,
Hudson
L. G.
(
1996
).
Enhanced modulation of keratinocyte motility by transforming growth factor-(TGF-) relative to epidermal growth factor (EGF).
J. Invest. Dermatol
106
,
590
597
Chakrabarty
S.
,
Rajagopal
S.
,
Huang
S.
(
1995
).
Expression of antisense epidermal growth factor receptor RNA downmodulates the malignant behavior of human colon cancer cells.
Clin. Exp. Metast
13
,
191
195
Chen
P.
,
Gupta
K.
,
Wells
A.
(
1994
).
Cell movement elicited by epidermal growth factor receptor requires kinase and autophosphorylation but is separable from mitogenesis.
J. Cell Biol
124
,
547
555
Chen
P.
,
Xie
H.
,
Sekar
M. C.
,
Gupta
K. B.
,
Wells
A.
(
1994
).
Epidermal growth factor receptor-mediated cell motility: phospholipase C activity is required, but MAP kinase activity is not sufficient for induced cell movement.
J. Cell Biol
127
,
847
857
Chen
P.
,
Murphy-Ullrich
J.
,
Wells
A.
(
1996
).
A role for gelsolin in actuating EGF receptor-mediated cell motility.
J. Cell Biol
134
,
689
698
Chen
Z.
,
Fisher
R. J.
,
Riggs
C. W.
,
Rhim
J. S.
,
Lautenberger
J. A.
(
1997
).
Inhibition of vascular endothelial growth factor-induced enothelial cell migration by ETS1 antisense oligonucleotides.
Cancer Res
57
,
2013
2019
Chicoine
M. R.
,
Silbergeld
D. L.
(
1995
).
The in vitro motility of human gliomas increases with increasing grade of malignancy.
Cancer
75
,
290
299
Diakonova
M.
,
Payrastre
B.
,
van Velzen
A. G.
,
Hage
W. J.
,
van Bergen en Henegouwen
P. M. P.
,
Boonstra
J.
,
Cremers
F. F. M.
,
Humbel
B. M.
(
1995
).
Epidermal growth factor induces rapid and transient association of phospholipase C-g1 with EGF-receptor and filamentous actin at membrane ruffles of A431 cells.
J. Cell Sci
108
,
2499
2509
DiMilla
P. A.
,
Stone
J. A.
,
Quinn
J. A.
,
Albelda
S. M.
,
Lauffenburger
D. A.
(
1993
).
Maximal migration of human smooth muscle cells on fibronectin and type IV collagen occurs at an intermediate attachment strength.
J. Cell Biol
122
,
729
737
Duband
J.-L.
,
Dufour
S.
,
Yamada
S.
,
Yamada
K.
,
Thiery
J.-P.
(
1991
).
Neural crest cell locomotin is induced by antibodies to1 integrins: A tool for studying the roles of substratum molecular avidity and density in migration.
J. Cell Sci
98
,
517
532
Dunn
G. A.
(
1983
).
Characterising a kinesis response: Time averaged measures of cell speed and directional persistence.
Agents and Actions [Suppl.]
12
,
14
33
Engebraaten
O.
,
Bjerkvig
R.
,
Pedersen
P. H.
,
Laerum
O. D.
(
1993
).
Effects of EGF, bFGF, NGF and PDGF(bb) on cell proliferative, migratory and invasive capacities of human brain-tumour biopsies in vitro.
Int. J. Cancer
53
,
209
214
Gail
M. H.
,
Boone
C. W.
(
1970
).
The locomotion of mouse fibroblasts in tissue culture.
Biophys. J
10
,
980
993
Goodman
S.
,
Risse
G.
,
von der Mark
K.
(
1989
).
The E8 subfragment of laminin promotes locomotion of myoblasts over extracellular matrix.
J. Cell Biol
109
,
799
809
Klemke
R. L.
,
Cai
S.
,
Giannini
A. L.
,
Gallagher
P. J.
,
DeLanerolle
P.
,
Cheresh
D. A.
(
1997
).
Regulation of cell motility by mitogen-activated protein kinase.
J. Cell Biol
137
,
481
492
Komuro
H.
,
Rakic
P.
(
1995
).
Dynamics of granule cell migration: A confocal microscopic study in acute cerebellar slice preparations.
J. Neurosci
15
,
1110
1120
Kundra
V.
,
Escobedo
J. A.
,
Kazlauskas
A.
,
Kim
H. K.
,
Rhee
S. G.
,
Williams
L. T.
,
Zetter
B. R.
(
1994
).
Regulation of chemotaxis by the platelet-derived growth factor receptor-.
Nature
367
,
474
476
Lauffenburger
D. A.
,
Horwitz
A. F.
(
1996
).
Cell migration: a physically integrated molecular process.
Cell
84
,
359
369
Li
X. B.
,
Nagayasu
H.
,
Hamada
J.
,
Hosokawa
M.
,
Takeichi
N.
(
1993
).
Enhancement of tumorigenicity and invasion capacity of rat mammary adenocarcinoma cells by epidermal growth factor and transforming growth factor-beta.
Jpn J. Cancer Res
84
,
1145
1149
Maldonado
B. A.
,
Furcht
L. T.
(
1995
).
Epidermal growth factor stimulates integrin-mediated cell migration of cultured human corneal epithelial cells on fibronectin and arginine-glycine-aspartic acid peptide.
Invest. Ophthalmol. Visual Sci
36
,
2120
2126
Matthay
M. A.
,
Thiery
J. P.
,
Lafont
F.
,
Stampfer
F.
,
Boyer
B.
(
1993
).
Transient effect of epidermal growth factor on the motility of an immortalized mammary epithelial cell line.
J. Cell Sci
106
,
869
878
Nelson
J.
,
Allen
W. E.
,
Scott
W. N.
,
Bailie
J. R.
,
Walker
B.
,
McFarren
N. V.
,
Wilson
D. J.
(
1995
).
Murine epidermal growth factor (EGF) fragment (33–42) inhibits both EGF-and laminin-dependent enothelial cell motility and angiogenesis.
Cancer Res
55
,
3772
3776
Othmer
H. G.
,
Dunbar
S. R.
,
Alt
W.
(
1988
).
Models of dispersal in biological systems.
J. Math. Biol
26
,
263
298
Palecek
S. P.
,
Loftus
J. C.
,
Ginsburg
M. H.
,
Lauffenburger
D. A.
,
Horwitz
A. F.
(
1997
).
Integrin-ligand binding properties govern cell migration speed through cell-substratum adhesiveness.
Nature
385
,
537
540
Pruss
R. M.
,
Herschman
H. R.
(
1977
).
Variants of 3T3 cells lacking mitogenic response to epidermal growth factor.
Proc. Nat. Acad. Sci. USA
74
,
3918
3921
Reddy
C. C.
,
Wells
A.
,
Lauffenburger
D.
(
1994
).
Proliferative response of fibroblasts expressing internalization-deficient EGF receptors is altered via differential EGF depletion effects.
Biotech. Progr
10
,
377
384
Ridley
A. J.
,
Hall
A.
(
1992
).
The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors.
Cell
70
,
389
399
Ridley
A. J.
,
Paterson
H. F.
,
Johnston
C. L.
,
Diekmann
D.
,
Hall
A.
(
1992
).
The small GTP binding protein rac regulates growth factor-induced membrane ruffling.
Cell
70
,
401
410
Ridley
A. J.
,
Comoglio
P. M.
,
Hall
A.
(
1995
).
Regulation of scatter factor/hepatocyte growth factor responses by ras, rac and rho in MDCK cells.
Mol. Cell Biol
15
,
1110
1122
Segall
J. E.
,
Tyerech
S.
,
Boselli
L.
,
Masseling
S.
,
Helft
J.
,
Chan
A.
,
Jones
J.
,
Condeelis
J.
(
1996
).
EGF stimulates lamellipod extension in metastatic mammary adenocarcinoma cells by an actin-dependent mechanism.
Clin. Exp. Metast
14
,
61
72
Siegal
G. P.
,
Wang
M.-H.
,
Rinehart
C. A.
,
Kennedy
J. W.
,
Goodly
L. J.
,
Miller
Y.
,
Kaufman
D. G.
,
Singh
R. K.
(
1993
).
Development of a novel human extracellular matrix for quantitation of the invasiveness of human cells.
Cancer Lett
69
,
123
132
Tessier-Lavigne
M.
,
Goodman
C. S.
(
1996
).
The molecular biology of axon guidance.
Science
274
,
1123
1133
Thommen
R.
,
Humar
R.
,
Misevic
G.
,
Pepper
M. S.
,
Hahn
A. W.
,
John
M.
,
Battegay
E. J.
(
1997
).
PDGF-BB increases endothelial migration on cord movements during angiogenesis in vitro.
J. Cell. Biochem
64
,
403
413
Turner
T.
,
Chen
P.
,
Goodly
L. J.
,
Wells
A.
(
1996
).
EGF receptor signaling enhances in vivo invasiveness of DU-145 human prostate carcinoma cells.
Clin. Exp. Metast
14
,
409
418
Ushiro
S.
,
Ono
M.
,
Izumi
H.
,
Kohno
K.
,
Taniguchi
N.
,
Higashiyama
S.
,
Kuwano
M.
(
1996
).
Heparin-binding epidermal growth factor-like growth factor: p91 activation induction of plasminogen activator/inhibitor, and tubular morphogenesis in human microvascular endothelial cells.
Jpn J. Cancer Res
87
,
68
77
Wells
A.
,
Bishop
J. M.
(
1988
).
Genetic determinants of neoplastic transformation by the retroviral oncogene v-erbB.
Proc. Nat. Acad. Sci. USA
85
,
7597
7601
Wells
A.
,
Welsh
J. B.
,
Lazar
C. S.
,
Wiley
H. S.
,
Gill
G. N.
,
Rosenfeld
M. G.
(
1990
).
Ligand-induced transformation by a non-internalizing EGF receptor.
Science
247
,
962
964
Welsh
J. B.
,
Gill
G. N.
,
Rosenfeld
M. G.
,
Wells
A.
(
1991
).
A negative feedback loop attenuates EGF-induced morphological changes.
J. Cell Biol
114
,
533
543
Wennstrom
S.
,
Siegbahn
A.
,
Yokote
K.
,
Arvidsson
A. K.
,
Heldin
C. H.
,
Mori
S.
,
Claesson-Welsh
L.
(
1994
).
Membrane ruffling and chemotaxis transduced by the PBDF-receptor require the binding site for phosphatidylinositol 3 kinase.
Oncogene
9
,
651
660
Wijelath
E. S.
,
Carlsen
B.
,
Cole
T.
,
Chen
J.
,
Kothari
S.
,
Hammond
W. P.
(
1997
).
Oncostatin M induces basic fibroblast growth factor expression in endothelial cells and promotes endothelial cell proliferation, migration and spindle morphology.
J. Cell Sci
110
,
871
879
Wilson
S. E.
,
He
Y. G.
,
Weng
J.
,
Zieske
J. D.
,
Jester
J. V.
,
Schultz
G. S.
(
1994
).
Effect of epidermal growth factor, hepatocyte growth factor, and keratinocyte growth factor, on proliferation, motility, and differentiation of human corneal epithelial cells.
Exp. Eye Res
59
,
665
678
Xie
H.
,
Turner
T.
,
Wang
M.-H.
,
Singh
R. K.
,
Siegal
G. P.
,
Wells
A.
(
1995
).
In vitro invasiveness of DU-145 human prostate carcinoma cells is modulated by EGF receptor-mediated signals.
Clin. Exp. Metast
13
,
407
419
Xie
H.
,
Pallero
M. A.
,
Gupta
D.
,
Chang
P.
,
Ware
M. F.
,
Witke
W.
,
Kwiatkowski
D. J.
,
Lauffenburger
D. A.
,
Murphy-Ullrich
J. E.
,
Wells
A.
(
1998
).
EGF receptor regulation of cell motility: EGF induces disassembly of focal adhesions independently of the motility-associated PLCsignaling pathway.
J. Cell Sci
111
,
615
624
This content is only available via PDF.