Hydrogen Peroxide Suppresses U937 Cell Death by Two Different Mechanisms Depending on Its Concentration

doi:10.1006/excr.1999.4409

Experimental Cell Research

Volume 248, Issue 2, 1 May 1999, Pages 430-438

https://doi.org/10.1006/excr.1999.4409 Get rights and content

Abstract

To investigate the mechanisms of H₂O₂adaptation in mammalian cells, we exposed human U937 leukemia cells to 0.05 mM H₂O₂. This treatment significantly suppressed cell death and DNA fragmentation induced by a subsequent challenge with 1 mM H₂O₂. A more dramatic protection was observed when cells were pretreated with 0.25 mM H₂O₂. Pretreatment with either 0.05 or 0.25 mM H₂O₂also imparted cells with a survival advantage against serum withdrawal and C₂-ceramide treatment. H₂O₂was found to be a mediator of cell death induced by serum withdrawal, but not by the addition of C₂-ceramide. Interestingly, 0.25 mM H₂O₂greatly induced glutathione peroxidase, a H₂O₂-consuming enzyme, whereas 0.05 mM H₂O₂did not. Consistent with observation, pretreatment with 0.25 mM H₂O₂resulted in a great reduction of cellular oxidant levels as determined by 2′7′-dichlorofluorescein fluorescence, and it also prevented elevation of oxidant levels upon subsequent challenge with 1 mM H₂O₂or with serum withdrawal. These effects were not observed in cells pretreated with 0.05 mM H₂O₂. The sum of the data indicated that H₂O₂suppresses cell death by two different mechanisms depending on its concentration: Relatively high concentrations enhance cellular antioxidant capacity, and lower concentrations block the lethal action of H₂O₂.

References (37)

S. Toyokuni et al.
Persistent oxidative stress in cancer
FEBS Lett.
(1995)
J.M.S. Davies et al.
Transient adaptation to oxidative stress in yeast
Arch. Biochem. Biophys.
(1995)
A.G. Wiese et al.
Transient adaptation to oxidative stress in mammalian cells
Arch. Biochem. Biophys.
(1995)
M.F. Christman et al.
Positive control of a regulon for defenses against oxidative stress and some heat shock proteins inSalmonella typhimurium
Cell
(1985)
S. Shull et al.
Differential regulation of antioxidant enzymes in response to oxidants
J. Biol. Chem.
(1991)
C. Nosseri et al.
Possible involvement of poly(ADP-ribosyl) polymerase in triggering stress-induced apoptosis
Exp. Cell Res
(1994)
D.M. Hockenbery et al.
Bcl-2 functions in an antioxidant pathway to prevent apoptosis
Cell
(1993)
N. Avissar et al.
Plasma selenium-dependent glutathione peroxidase
J. Biol. Chem.
(1989)
M. Maiorino et al.
Phospholipid hydroperoxide glutathione peroxidase is the 18-kDa selenoprotein expressed in human tumor cell line
J. Biol. Chem.
(1991)
F.-F. Chu et al.
Expression, characterization, and tissue distribution of a new cellular selenium-dependent glutathione peroxidase, GSHPx-GI
J. Biol. Chem.
(1993)

M.B. Hampton et al.

Dual regulation of caspase activity by hydrogen peroxide: implications for apoptosis

FEBS Lett.

(1997)

B.A. Freeman et al.

Biology of disease; free radicals and tissue injury

Lab. Invest.

(1982)

S.B. Farr et al.

Oxidative stress responses inEscherichia coliSalmonella typhimurium

Microbiol. Rev.

(1991)

B. Demple

Regulation of bacterial oxidative stress genes

Annu. Rev. Genet.

(1991)

J.T. Greenberg et al.

Positive control of a global antioxidant defense regulon activated by superoxide-generating agents inEscherichia coli

Proc. Natl. Acad. Sci. USA

(1990)

J. Wu et al.

Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon inEscherichia coli

J. Bacteriol.

(1991)

D. Lu et al.

Molecular adaptation of vascular endothelial cells to oxidative stress

Am. J. Physiol.

(1993)

D. Mangan et al.

Lipopolysaccharide, tumor necrosis factorα, and IL-1β prevent programmed cell death (apoptosis) in human peripheral blood monocytes

J. Immunol.

(1991)

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Regular ArticleHydrogen Peroxide Suppresses U937 Cell Death by Two Different Mechanisms Depending on Its Concentration

Abstract

FEBS Lett.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Cell

J. Biol. Chem.

Exp. Cell Res

Cell

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

FEBS Lett.

Biology of disease; free radicals and tissue injury

Lab. Invest.

Oxidative stress responses inEscherichia coliSalmonella typhimurium

Microbiol. Rev.

Regulation of bacterial oxidative stress genes

Annu. Rev. Genet.

Positive control of a global antioxidant defense regulon activated by superoxide-generating agents inEscherichia coli

Proc. Natl. Acad. Sci. USA

Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon inEscherichia coli

J. Bacteriol.

Molecular adaptation of vascular endothelial cells to oxidative stress

Am. J. Physiol.

Lipopolysaccharide, tumor necrosis factorα, and IL-1β prevent programmed cell death (apoptosis) in human peripheral blood monocytes

J. Immunol.

Regular Article
Hydrogen Peroxide Suppresses U937 Cell Death by Two Different Mechanisms Depending on Its Concentration