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Role of the Alkali Labile Sites, Reactive Oxygen Species and Antioxidants in DNA Damage Induced by Methylated Trivalent Metabolites of Inorganic Arsenic

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Abstract

In the last decade arsenic metabolism has become an important matter of discussion. Methylation of inorganic arsenic (iAs) to monomethylarsonic acid (MMAV) and dimethylarsinic acid (DMAV) is considered to decrease arsenic toxicity. However, in addition to these pentavalent metabolites, the trivalent metabolites monomethylarsonous (MMAIII) and dimethylarsinous acid (DMAIII) have been identified recently as intermediates in the metabolic pathway of arsenic in cultured human cells. To examine the role of oxidative damage in the generation of DNA strand breaks by methylated trivalent arsenic metabolites, we treated human lymphocytes with both metabolites at non-cytotoxic concentrations. We further tested whether these effects are sensitive to modulation by the antioxidants ascorbate (Vitamin C) and selenomethionine (Se-Met). Both trivalent metabolites produced oxidative stress related DNA damage, consisting of single strand breaks and alkali-labile sites, with MMAIII being more potent at low concentrations than DMAIII. Neither MMAIII nor DMAIII induced DNA-double strand breaks. The oxidative stress response profiles of the metabolites were parallel as determined by lipid peroxidation induction. MMAIII induced peroxidation from the lowest concentration tested, while effects of DMAIII were apparent only at concentrations above 10 μM. The antioxidant Se-Met exhibited a more pronounced inhibition of trivalent arsenic metabolite-induced oxidative-DNA damage than did vitamin C. The present findings suggest that DNA damage by methylated trivalent metabolites at non-cytotoxic concentrations may be mediated by a mix of reactive oxygen and nitrogen oxidized species.

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Abbreviations

iAs:

inorganic arsenic

iAsIII :

arsenite

MMAIII :

monomethylarsonous acid

MMAV :

monomethylarsonic acid

DMAIII :

dimethylarsinous acid

DMAV :

dimethylarsinic acid Se-Met- selenomethionine

ALS:

alkali labile sites

LMPA:

low melting point agarose

Et-Br:

ethidium bromide

Tris:

Tris[hydroxymethyl]aminomethane

DMSO:

dimethyl sulfoxide

FDA:

5,6-carboxiacetate of fluoresceine

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Soto-Reyes, E., Del Razo, L.M., Valverde, M. et al. Role of the Alkali Labile Sites, Reactive Oxygen Species and Antioxidants in DNA Damage Induced by Methylated Trivalent Metabolites of Inorganic Arsenic. Biometals 18, 493–506 (2005). https://doi.org/10.1007/s10534-005-0858-7

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