Folic acid protects SWV/Fnn embryo fibroblasts against arsenic toxicity.

It has been proposed that arsenic exerts its toxic effects, in part, by perturbing cellular methyl metabolism. Based on the hypothesis that folic acid treatment will attenuate the cytotoxic and growth inhibitory effects of arsenic, SWV/Fnn embryo fibroblasts were cultured in media supplemented with various concentrations of folic acid during treatment with sodium arsenite or dimethylarsinic acid (DMA). It was found that folic acid protects SWV/Fnn embryo fibroblasts from sodium arsenite and DMA cytotoxicity in a dose-dependent manner. In contrast, folic acid supplementation has no effect on toxicity resulting from treatment with ethanol or staurosporine, suggesting that folic acid is not generally protective against necrosis and apoptosis. Although folic acid protects against acute arsenic toxicity, this agent shows a modest and delayed ability to attenuate the growth inhibitory effect of arsenic on these cells. These results support a model in which perturbations of methyl metabolism contribute to the acute cytotoxicity of arsenic.

Sodium arsenite inhibits terminal differentiation of murine C3H 10T1/2 preadipocytes.

Cancer represents an imbalance between cell proliferation and differentiation, two processes that are coordinately and antagonistically regulated. Aberrant cell proliferation is considered to be an important etiological factor in the development of arsenic-induced cancer, suggesting that arsenic also dysregulates differentiation. Based on evidence that arsenic modulates mitogenic events that antagonize the process of differentiation, this study addresses the hypothesis that sodium arsenite inhibits insulin/dexamethasone-induced differentiation of C3H 10T1/2 preadipocytes; it was further postulated that arsenic-treated cells retain mitogenic responsiveness under differentiating conditions. To test this hypothesis, the differentiation capacity of C3H 10T1/2 preadipocytes was examined in control cells and cells treated with sodium arsenite. Differentiation was assessed morphologically and quantified by Oil Red-O staining of accumulated lipids. The effect of long-term arsenic exposure on mitogenic competence was quantified by flow cytometry, [(3)H]thymidine incorporation, and cell counting under conditions favorable for adipocyte differentiation. Results indicate that arsenic inhibits morphological differentiation of wild-type C3H 10T1/2 preadipocytes. Short-term arsenic exposure inhibits differentiation in a dose-dependent manner, with arsenic concentrations > or = 3 microM producing a significant inhibition of dexamethasone/insulin-induced lipid accumulation. Furthermore, arsenic-treated cells exhibit an accentuated response to mitogenic stimulation under differentiating conditions. These data suggest that arsenic exposure results in the inhibition of cellular programming required for terminal differentiation of C3H 10T1/2 preadipocytes and that cells acquire mitogenic hyperresponsiveness. The ability of arsenic to dysregulate the balance between proliferation and differentiation is proposed to be one mechanism by which this metalloid causes cancer in humans.

Sodium arsenite-induced dysregulation of proteins involved in proliferative signaling.

It is well accepted that arsenic is a human carcinogen, yet its mechanism of action is not defined. Arsenic cannot be classified as an initiating agent or as a promoter, although altered proliferative responsiveness has been proposed as a mechanism by which arsenic exerts its carcinogenic effects. Based on the hypothesis that arsenic exposure results in modulation of both positive and negative regulators of cell proliferation, this study examined physiological and biochemical changes in the proliferative response of murine fibroblasts grown long-term in the maximum tolerated concentration of sodium arsenite. In response to EGF stimulation, DNA synthesis and the proportion of cells entering S phase of the cell cycle both were increased in cells grown long-term in arsenic compared to control cells. Analysis of positive proliferative regulators revealed an increase in the expression of c-myc and E2F-1, thereby supporting the hypothesis that arsenic increases activity of positive growth modulators. In contrast, the activity and expression of ERK-2 were unchanged, as was the expression of EGF-receptor and mSOS. When negative regulators of proliferation were examined, expression levels of MAP kinase phosphatase-1 and p27(Kip1) were found to be lower in arsenic-treated cells compared to control cells; this result supports a model in which arsenic disinhibits normal regulation of cell proliferation. Taken together, these data indicate that long-term exposure to sodium arsenite creates conditions within the cell consistent with sensitization to mitogenic stimulation. It is further postulated that the observed changes in mitogenic signaling proteins contribute to the carcinogenic property of arsenic.