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J. Biol. Chem., Vol. 278, Issue 10, 8058-8064, March 7, 2003
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From the Laboratory of Pharmacology and Chemistry, NIEHS, National
Institutes of Health,
Research Triangle Park, North Carolina 27709
We have investigated the role played by GSH
efflux in apoptosis of human HaCaT keratinocytes induced by UVA
irradiation. UVA irradiation of HaCaT cells caused a rapid rise in GSH
efflux across the intact cell membrane, followed by an increase in
apoptosis. GSH efflux was stimulated by glucose and was reduced by the
addition of exogenous GSH and intracellular GSH depletion by buthionine sulfoximine, suggesting that GSH transport is active and is influenced by the GSH concentration gradient across the cell membrane. Verapamil and cyclosporin A, blockers of the multidrug resistance-associated protein, decreased UVA-induced GSH efflux. GSH efflux occurred within
2 h of UVA irradiation, suggesting that the stimulation of GSH
efflux is due to an increase in the activity of pre-existing multidrug
resistance-associated protein transporter carrier. Although inhibition
of GSH efflux did not affect caspase activation and DNA fragmentation,
it delayed the gradual increase in plasma membrane permeability and
reduced phosphatidylserine translocation in HaCaT cells. It is
therefore likely that upon UVA irradiation, GSH efflux increased the
intracellular oxidative stress without intervention of reactive oxygen
species, thus resulting in more phosphatidylserine externalization and
membrane rearrangement. These provide targets for macrophage
recognition and phagocytosis and thus minimize the potential to invoke
inflammation or neoplastic transformation.
Role of Reduced Glutathione Efflux in Apoptosis of Immortalized
Human Keratinocytes Induced by UVA*
,
*
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.: 919-541-4751;
Fax: 919-541-5750; E-mail: he3@niehs.nih.gov.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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