Cellular uptake and metabolic reduction of pentavalent to trivalent arsenic as determinants of cytotoxicity and morphological transformation.
Cytotoxicity, morphological neoplastic transformation, cellular uptake and metabolic reduction were determined in BALB/3T3 Cl A31-1-1 cells for trivalent arsenic (sodium arsenite, As3+) and for pentavalent arsenic (sodium arsenate, As5+). The levels of cellular uptake of 73As-labelled sodium arsenite and arsenate were dose-dependent and highest in the first hour. At equimolar concentration (3 X 10(-6) M), cellular uptake was 4-fold higher for As3+ than for As5+. Cytotoxicity was higher for As3+ than for As5+, but when correlated to total As cell burden it showed no significant difference for the two forms. Morphological transformation focus assays showed transforming activity for both As3+ and As5+, with relative transformation frequencies also of approximately 4:1. Recovery from the cytosol after exposure for 1-24 h was greater than 90% for either form of absorbed As. Exposure to As3+ yielded 100% as As3+ in cytosol, but exposure to As5+ yielded greater than 70% as As3+, showing a high rate of intracellular metabolic reduction. No methylated metabolites were detected by ion-exchange chromatography. After 24-h incubation in cell-free medium, oxidation of As3+ to As5+ occurred up to 30% of the dose, but incubation in the presence of cells lowered the oxidation level to 4%. As5+ was recovered unchanged from cell-free medium (24-h incubation), but in the presence of the cells it yielded up to 5% as As3+ within 24 h and the cumulative release of As3+ by cells exposed to As5+ was dose-dependent. Glutathione depletion by diethylmaleate inhibited reduction of As5+ to As3+ by these cells up to 25% of controls, showing that As5+ reduction is partly dependent on glutathione. These results suggest that As3+ is the form responsible for the cytotoxic and transforming effects, independently of the valence state of the inorganic arsenic in the culture medium.
D000818:Animals; D001149:Arsenates / Q000378:metabolism / Q000633:toxicity*; D001151:Arsenic / Q000378:metabolism / Q000633:toxicity*; D018053:Arsenites; D001692:Biological Transport; D001711:Biotransformation; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D017670:Sodium Compounds
D001149:Arsenates; D018053:Arsenites; D017670:Sodium Compounds; C017947:sodium arsenite; C009277:sodium arsenate; D001151:Arsenic