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Enhanced glutathione S-transferase expression in 2-hydroxyamino- 1-methyl-6-phenylimidazo[4,5-b]pyridine-resistant IEC-18 cells

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Abstract

In the present study we show that repeated exposure of the rat intestinal epithelial cell line IEC-18 to 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-OH-PhIP), from a toxicological point of view the most relevant phase-1 metabolite of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP, the main heterocyclic aromatic amine present in processed meat), led to the selection of N-OH-PhIP-resistant IEC-18 cells. This phenomenon was accompanied by a fivefold increase in total glutathione S-transferase (GST) activity, measured with the broad-spectrum substrate 1-chloro-2,4-dinitrobenzene, in the N-OH-PhIP-resistant IEC-18 cells. Furthermore, a Western blotting analysis revealed that the expression of GST subunits A1, A3, A4, M1 and P1 was enhanced in the N-OH-PhIP-resistant IEC-18 cells.

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Abbreviations

CDNB:

1-chloro-2,4-dinitrobenzene

CYP:

cytochrome P450

DMSO:

dimethyl sulfoxide

GST:

glutathione S-transferase

HAA:

heterocyclic aromatic amine

MTT:

3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl tetrazolium bromide

N-OH-PhIP:

2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine

NAT:

N-acetyltransferase

PhIP:

2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine

RT-PCR:

reverse transcription–polymerase chain reaction

SULT:

sulfotransferase

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Authors and Affiliations

  1. Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany

    W. Teubner, J. I. Fuchs & P. Steinberg

  2. Chair of Nutritional Toxicology, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany

    P. Steinberg

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  1. W. Teubner
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  2. J. I. Fuchs
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Correspondence to P. Steinberg.

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Teubner, W., Fuchs, J.I. & Steinberg, P. Enhanced glutathione S-transferase expression in 2-hydroxyamino- 1-methyl-6-phenylimidazo[4,5-b]pyridine-resistant IEC-18 cells. Cell Biol Toxicol 23, 153–161 (2007). https://doi.org/10.1007/s10565-006-0094-0

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