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J. Biol. Chem., Vol. 277, Issue 22, 19402-19407, May 31, 2002
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From the Free Radical Research Group, Department of Pathology,
Christchurch School of Medicine and Health Sciences, P. O. Box 4345, Christchurch, New Zealand
Diphenyleneiodonium (DPI) is a broad-spectrum
flavoprotein inhibitor commonly used to inhibit oxidant production by
the NADPH oxidase of phagocytic and nonphagocytic cells. A previous
study has shown that DPI can sensitize T24 bladder carcinoma cells to Fas-mediated apoptosis. We observed DPI to deplete intracellular reduced glutathione (GSH) in T24 cells and a range of other primary and
transformed cell types. The effect was immediate, with 50% loss of
intracellular GSH within 2 h of treatment with DPI. The glutathione was quantitatively recovered in the extracellular medium,
indicating that efflux was occurring. The loss of GSH was blocked with
bromosulfophthalein, an inhibitor of the canalicular GSH transporters.
We conclude that DPI induces a dramatic efflux of cellular GSH from T24
cells via a specific transport channel. This provides a
potential mechanism for its proapoptotic effect, and it also has
important implications for the regulation of glutathione homeostasis in cells.
Diphenyleneiodonium Triggers the Efflux of Glutathione from
Cultured Cells*
and
*
This work was supported in part by an Otago University
Research Grant and by the Cancer Society of New Zealand.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 64-3-364-0565;
Fax: 64-3-364-1083; E-mail: juliet.pullar@chmeds.ac.nz.
§
Supported by a grant from the Marsden Fund of the Royal Society of
New Zealand.
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