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J. Biol. Chem., Vol. 266, Issue 34, 22968-22974, 12, 1991
JA Shayman, GD Deshmukh, S Mahdiyoun, TP Thomas, D Wu, FS Barcelon and NS Radin
Two independent approaches were employed to explore the potential role of
endogenous glucosylceramide or a closely related glucosphingolipid in
mediating the cellular proliferation of Madin-Darby canine kidney cells.
First, cultured cells were depleted of glucosphingolipids by exposure to a
glucosylceramide synthase inhibitor, D-threo-1-phenyl-2-
decanoylamino-3-morpholino-1-propanol. This agent markedly inhibited cell
growth and DNA synthesis in a time- and concentration-dependent manner.
Second, cells were grown in the presence of conduritol B epoxide, an
inhibitor of glucosylceramide beta-D-glucosidase. Exposure of cells to this
inhibitor resulted in the time-dependent accumulation of glucosylceramide
with a corresponding increase in cellular proliferation. Alterations in
protein kinase C activity were evaluated as a potential mechanism for these
effects on growth. Both membrane- and cytosol-associated protein kinase C
(PKC) activity declined under conditions of glucosylceramide synthase
inhibition and increased under conditions of beta-glucosidase inhibition.
The changes in PKC activity were evident after DEAE-cellulose purification.
Diacylglycerol levels increased in response to both glucosylceramide
synthase and beta- glucosidase inhibition. Ceramide and sphingosine levels
changed only in the presence of
D-threo-1-phenyl-2-decanoylamino-3-morpholino-1- propanol, increasing due
to lack of conversion to glucosylceramide. However, the elevation in
endogenous sphingosine was probably insufficient to account for the
decrease in PKC, considering the high level of diacylglycerol in the cells.
These data demonstrate an association between glucosylceramide levels, PKC
activity, and cell growth.
Modulation of renal epithelial cell growth by glucosylceramide. Association with protein kinase C, sphingosine, and diacylglycerol
Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor 48109.
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