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Mutagenicity of the mycotoxin patulin in cultured Chinese hamster V79 cells, and its modulation by intracellular glutathione

  • Genotoxicity
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Abstract

Because the ability of the mycotoxin patulin (PAT) to cause gene mutations in mammalian cells is still ambiguous, we have studied the mutagenicity of PAT at the hypoxanthine–guanine phosphoribosyltransferase (HPRT) gene locus in cultured Chinese hamster V79 cells with normal, depleted, and elevated glutathione (GSH) levels. PAT was more toxic to GSH-depleted cells than to normal cells and caused an increase of the intracellular GSH level in normal and GSH-depleted cells. It also caused synchronization of the cell cycle due to a temporary accumulation of cells in the G2/M phase; this G2/M arrest was more persistent in GSH-depleted than in normal cells. PAT gave rise to a clear and concentration-dependent induction of HPRT mutations at non-cytotoxic concentrations in V79 cells with normal GSH level; the lowest PAT concentration causing a significant number of mutant cells was 0.3 micromolar, and the mutagenic potency of PAT equaled that of the established mutagen 4-nitroquinoline-N-oxide. The mutagenicity of PAT was again more pronounced, by a factor of about three, in GSH-depleted V79 cells. Elevated GSH levels abolished all observed effects of PAT. These data support the notion that PAT is a mutagenic mycotoxin, in particular in cells with low GSH concentration. The ability of PAT to cause gene mutations in mammalian cells might have a bearing on its carcinogenicity.

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Abbreviations

ATM:

Ataxia telangiectasia mutated kinase

ATR:

ATM-related kinase

BSO:

Buthionine sulfoximine

Cdk:

Cyclin-dependent kinase

DMEM:

Dulbecco’s modified Eagle medium

DMSO:

Dimethyl sulfoxide

DTNB:

5,5′-Dithiobis-(2-nitrobenzoic acid)

EDTA:

Ethylenediaminetetraacetic acid

FCS:

Fetal calf serum

GSH:

Glutathione

GSH-EE:

Glutathione ethyl ester

HPRT:

Hypoxanthine-guanine phosphoribosyltransferase

MClB:

Monochlorobimane

NQO:

4-Nitroquinoline-N-oxide

PAT:

Patulin, 4-hydroxy-4H-furo(3,2c)pyran-2(6H)-one

PBS-CMF:

Phosphate-buffered saline, calcium- and magnesium-free

6-TG:

6-Thioguanine

UV:

Ultraviolet

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Acknowledgements

We thank Ute Franz for helping with the studies on the cell-cycle distribution, Aleka Vavelidou for microscopic analysis of the membrane integrity, and Jörg Wagner and Lars Rohnstock for their help with the HPRT assay. This study has been supported by the Deutsche Forschungsgemeinschaft (Grant Me 574/14–2).

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  1. Institute of Food Chemistry and Toxicology, University of Karlsruhe, P.O. Box 6980, 76128, Karlsruhe, Germany

    David M. Schumacher, Manfred Metzler & Leane Lehmann

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  1. David M. Schumacher
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  2. Manfred Metzler
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Correspondence to Leane Lehmann.

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Schumacher, D.M., Metzler, M. & Lehmann, L. Mutagenicity of the mycotoxin patulin in cultured Chinese hamster V79 cells, and its modulation by intracellular glutathione. Arch Toxicol 79, 110–121 (2005). https://doi.org/10.1007/s00204-004-0612-x

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