Low-level self-tolerance to arsenite in human HepG2 cells is associated with a depressed induction of micronuclei.

Arsenic (As) is one of the most important global environmental toxicants. There is evidence that humans may develop tolerance to As's toxicity. For instance, it is known that uptake of small amounts of As leads to an acquisition of elevated resistance to the element's acute toxicity. Moreover, it was suggested that As-exposed native Andean females of Atacameño ethnicity may have acquired resistance to skin cancer. It is not known how such adaptation could be mechanistically conferred. In this context, the biological selection and cloning of human cells tolerant to As provides a valuable approach to investigate this question. By the means of a 12 weeks culture with increasing doses of As, three different As-resistant clones of the human hepatoma cell line HepG2 were selected. These three clones were similarly and roughly two-fold resistant to the acute toxicity of arsenite (50% reduction of neutral red (NR) uptake at 65 microM versus 115 microM; HepG2 control versus clones HepG2 K1, HepG2 K11 and HepG2 K14, respectively). Moreover, in the cytokinesis-block micronucleus test, these clones showed a significantly reduced induction of micronuclei (MNi) indicating elevated resistance to As genotoxicity as well (e.g. mean MNi rates at a concentration of 25 microM arsenite: 28.5 (control) versus 21.6 (HepG2 K1), 18 (HepG2 K11), and 16 (HepG2 K14), respectively, each P<0.05). The tolerance was neither associated with mRNA induction of putatively As-extruding membrane transporters multidrug resistance-associated protein 1 (MRP1), 2, or 3 nor to mRNA induction of the ubiquitously expressed mammalian ABC half-transporter UMAT (ABCB6). Changes in the metabolic methylation of As could not be detected. There were no differences in the cellular levels of GSH when comparing the clones and the parental line. Taken together the data showed that low-level tolerance to As-mediated cytotoxicity in human HepG2 cells was associated with enhanced resistance to As-induced DNA damage as well.