Antagonism of cadmium cytotoxicity by differentiation inducers

Studies on the antagonism of toxicity can provide information about toxic mechanisms and suggest chemotherapeutic strategies. A rapid cell growth assay that measures the effects of test agents on the accumulation of cell protein (Shopsis and Eng, Toxicol. Lett. 1985;26:1) has been applied to studies of the antagonism of the cytotoxicity of cadmium. Exposure of Balb/c mouse 3T3 cells to 15 mumol/L Cd2+ for 24 h or 7 mumol/L Cd2+ for 48 h caused a 50% decrease in total cell protein. Zn2+ and selenite ion, antagonists of Cd toxicity in vivo, antagonized Cd2+ cytotoxicity when added in micromolar concentrations at the initiation of exposure to Cd2+. A diverse group of chemicals that can induce differentiation in vitro in cultured erythroleukemia and other cells were also found to antagonize the cytotoxic effects of Cd2+ to 3T3 cells. Dimethyl sulfoxide (DMSO), hexamethylene bisacetamide, N,N-dimethyl formamide, N-methyl formamide, dimethyl acetamide, hypoxanthine, hemin, ouabain, and sodium butyrate, when added to cultures simultaneously with Cd2+, each antagonized Cd2+ toxicity. These agents were used at concentrations equal to or lower than the concentrations at which they induce cellular differentiation. Other cytotoxicity assays and morphological studies confirmed these observations. DMSO added as much as 6 h after the initiation of a 24-h exposure to Cd2+ still protected cells; conversely, pretreatment of cultures with butyrate or DMSO for 24 h followed by their removal did not confer protection against subsequent Cd2+ challenge. Ethanol and methanol (noninducers of differentiation) did not antagonize Cd2+ cytotoxicity, and differentiation-inducing agents did not protect the cells from Zn(2+)- or Hg(2+)-induced cytotoxicity. DMSO treatment does not induce an increase in the concentrations of metallothionein or glutathione in these cells.