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(Received for publication, June 20, 1996, and in revised form, August 7, 1996)
From the Department of Medicine, University of California, San
Diego, School of Medicine, San Diego, California 92103
Epidermal growth factor (EGF) and carbachol both
inhibit calcium-activated chloride secretion by the human colonic
epithelial cell line, T84. Although the inhibitory
mechanism for the carbachol effect involves the 3,4,5,6-isomer of
inositol tetrakisphosphate, the mechanisms responsible for the EGF
effect have not yet been fully elucidated. Here, we studied the role of
phosphatidylinositol 3-kinase (PI 3-kinase) in the inhibitory
effect of EGF. The PI 3-kinase inhibitor, wortmannin, slightly
increased basal chloride secretion and potentiated the secretory
response to thapsigargin. Wortmannin also partially reversed
EGF-induced, but not carbachol-induced, inhibition of
thapsigargin-stimulated chloride secretion. Wortmannin alone had no
effect on carbachol- or histamine-induced chloride secretion and
completely reversed EGF-induced inhibition of the secretory response to
these agonists. EGF, carbachol, histamine, and thapsigargin all
increased levels of the 85-kDa regulatory subunit of PI 3-kinase in
antiphosphotyrosine immunoprecipitates. However, only EGF
significantly increased levels of the 110-kDa catalytic
subunit. Furthermore, only EGF increased PI 3-kinase activity in an
in vitro kinase assay. High levels of
phosphatidylinositol ()-monophosphate were present in unstimulated
cells and significantly reduced by wortmannin. EGF, but not carbachol,
rapidly increased levels of phosphatidylinositol (3,4)-bisphosphate
and phosphatidylinositol (3,4,5)-trisphosphate. Production of these
lipids was also sensitive to wortmannin. Our data suggest that EGF
activates PI 3-kinase and that its lipid products may mediate the
inhibitory effect of EGF on calcium-dependent chloride
secretion. Our data also suggest that a phosphatidylinositol-specific
3-kinase activity is present in unstimulated T84 cells and
may regulate production of phosphatidylinositol ()-monophosphate and
basal secretory tone.
Volume 271, Number 43,
Issue of October 25, 1996
pp. 26588-26595
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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