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Vol. 9, Issue 8, 2185-2200, August 1998
Ludwig Institute for Cancer Research, University College London
Branch, London W1P 8BT, United Kingdom; and Department of Biochemistry
and Molecular Biology, University College London, London WC1E 6BT,
United Kingdom
Hepatocyte growth factor/scatter factor (HGF/SF) stimulates the
motility of epithelial cells, initially inducing centrifugal spreading
of colonies followed by disruption of cell-cell junctions and
subsequent cell scattering. In Madin-Darby canine kidney cells, HGF/SF-induced motility involves actin reorganization mediated by Ras,
but whether Ras and downstream signals regulate the breakdown of
intercellular adhesions has not been established. Both HGF/SF and
V12Ras induced the loss of the adherens junction proteins E-cadherin
and 
-catenin from intercellular junctions during cell spreading, and
the HGF/SF response was blocked by dominant-negative N17Ras. Desmosomes
and tight junctions were regulated separately from adherens junctions,
because they were not disrupted by V12Ras. MAP kinase,
phosphatidylinositide 3-kinase (PI 3-kinase), and Rac were required
downstream of Ras, because loss of adherens junctions was blocked by
the inhibitors PD098059 and LY294002 or by dominant-inhibitory mutants
of MAP kinase kinase 1 or Rac1. All of these inhibitors also
prevented HGF/SF-induced cell scattering. Interestingly, activated Raf
or the activated p110
subunit of PI 3-kinase alone did not induce
disruption of adherens junctions. These results indicate that
activation of both MAP kinase and PI 3-kinase by Ras is required for
adherens junction disassembly and that this is essential for the motile
response to HGF/SF.
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