Evidence for Free Radical Involvement in the Toxicity and Carcinogenicity of Chromate Dusts

"INTRODUCTIONEpidemiologic studies of workers in chromate-ore related industries, 1-3 and stainless steel welding and related occupations4 have shown that they have about 20-40 times higher risk of throat or respiratory track cancer than controls. Although the actual carcinogenic substances were not identified in these statistical studies, Cr(VI) compounds (for example, calcium chromate, zinc chromate, and lead chromate) were implicated as the causative agents, whereas Cr(III) compounds were not suspected as carcinogens. s These suggestions were supporte4 by laboratory studies wherein many Cr(VI) compounds produced sarcomas at the implant or injection sites.1,5 Squamous cell carcinomas and adenocarcinomas closely resembling human lung cancer were induced by intrabronchial implants of calcium chromate in rats, 6·7 whereas Cr(DI) oxide and Cr(lll) sulfate did not induce any tumor formation.6,7While the mechanism of the chromate-induced carcinogenicity is not fully understood, it is generally thought8 that it involves some damage to DNA. Specifically, it has been reported that: a) the chromate ion, henceforth referred to as chromate, can pass through the cell membrane and enter the cell while CR(DI) does not, 9 b) chromate does not interact with either native or denatured DNA 10 while Cr(III) does, 11 and c) the final Cr-DNA complex isolated from cellular reactions of chromate is Cr(III)-DNA, with Cr(III) binding to the phosphate groups.12,13 Thus the important question is: since Cr(DI) cannot pass through the membranes, how does Cr(III)-DNA complex form? For this to happen, Cr(VI) must be reduced to its lower oxidation states, 14 ultimately to Cr(III), by some reductants in the cellular environment. Unless this reduction occurs, the DNA would not be damaged and therefore no chromate carcinogenicity would ensue. Thus the reduction of chromate to its lower oxidation states seems to be a key step in the chromate carcinogenicity .1s One of the major reductants in cellular environments is thought to be glutathione (GSH), both outside and inside the cells.16-lB Some evidence for the role of GSH in the chromate toxicity was provided by recent studies showing that exposure of hamster cells to non-toxic levels of added selemite increases the levels of GSH as well as the Cr(VI)-induced DNA strand breaks,19 and that such DNA strand breaks in hepatocytes also change in direct proportion to the GSH content. 20,21 These observations were interpreted as implying that the reduction of chromate by GSH to some reactive intennediate is an important step in the chromate carcinogenicity. 20.21 In · the present undenaking22 we have used electron spin resonance (ESR) and spin-trap methodology to investigate the reduction of chromate by GSH and find evidence for the involvement of the glutathionyl radical (GS ·) as well as Cr(V)-intermediates."