DNA damage induced by phorbol ester-stimulated neutrophils is augmented by extracellular cofactors. Role of histidine and metals

Emily Shacter; Rosa L Lopez; Edward J Beecham; Siegfried Janz

doi:10.1016/S0021-9258(19)39206-3

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DNA damage induced by phorbol ester-stimulated neutrophils is augmented by extracellular cofactors. Role of histidine and metals

Journal article

Open access

Peer reviewed

DNA damage induced by phorbol ester-stimulated neutrophils is augmented by extracellular cofactors. Role of histidine and metals

Emily Shacter, Rosa L Lopez, Edward J Beecham and Siegfried Janz

The Journal of biological chemistry, Vol.265(12), pp.6693-6699

04/25/1990

DOI: 10.1016/S0021-9258(19)39206-3

PMID: 2157707

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Abstract

Activated neutrophils cause extensive DNA damage in neighboring nonphagocytic cells. To determine whether compounds in the extracellular milieu participate in the DNA damage process, murine neutrophils were cocultivated with target tumor cells in media of varying composition. Using the alkaline elution assay, it was found that the level of strand breaks induced was significantly higher (2.8-fold) in complex cell culture media than in minimal phosphate-buffered saline. Addition of amino acids in general and of histidine in particular increased the level of damage nearly to that observed in complete media (2.7- and 2.1-fold, respectively). The histidine stimulation was concentration-dependent and reached a maximum at 100-400 microM. The mechanism whereby this occurred is not proven but probably derived from chelation of metals and participation in a site-specific Fenton reaction. Addition of the cell-impermeable chelator EDTA dramatically inhibited induction of strand breaks by neutrophils in complete media and prevented the enhancement of damage induced by histidine in phosphate-buffered saline. None of the effects on neutrophil-induced damage could be attributed to modulation of the oxidative burst activity of the cells (O2- and H2O2 production). Histidine also enhanced induction of strand breaks by reagent H2O2. However, EDTA had no effect or actually increased the level of damage induced by both a bolus of H2O2 and a flux of H2O2 generated by glucose oxidase. The cell-permeable chelator o-phenanthroline inhibited both neutrophil- and H2O2-induced damage. The results indicate that secondary reactions involving extracellular amino acids and metals contribute significantly to neutrophil-induced DNA damage to neighboring cells. Moreover, the data show that the mechanism whereby neutrophils induce this damage cannot be attributed solely to secretion of H2O2.

Cell Line

Histidine - pharmacology

Tetradecanoylphorbol Acetate - pharmacology

Neutrophils - drug effects

Tumor Cells, Cultured - drug effects

Cells, Cultured

Neutrophils - physiology

Tumor Cells, Cultured - metabolism

Amino Acids - pharmacology

Hydrogen Peroxide - metabolism

Animals

Edetic Acid - pharmacology

Superoxides - metabolism

Mice

Mice, Inbred BALB C

DNA Damage

Kinetics

Plasmacytoma

Leukemia L1210

Neutrophils - metabolism

Hydrogen-Ion Concentration

Metals - pharmacology

Details

Title: Subtitle: DNA damage induced by phorbol ester-stimulated neutrophils is augmented by extracellular cofactors. Role of histidine and metals
Creators: Emily Shacter - Laboratory of Genetics, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892
Rosa L Lopez
Edward J Beecham
Siegfried Janz
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.265(12), pp.6693-6699
DOI: 10.1016/S0021-9258(19)39206-3
PMID: 2157707
ISSN: 0021-9258
eISSN: 1083-351X
Language: English
Date published: 04/25/1990
Academic Unit: Pathology
Record Identifier: 9984083869402771

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