Formation of diethylstilbestrol–DNA adducts in human breast epithelial cells and inhibition by resveratrol

Extensive evidence exists that the reaction of estrogen metabolites with DNA produces depurinating adducts that, in turn, induce mutations and cellular transformation. While it is clear that these estrogen metabolites result in a neoplastic phenotype in vitro, further evidence supporting the link between estrogen–DNA adduct formation and its role in neoplasia induction in vivo would strengthen the evidence for a genotoxic mechanism.Diethylstilbestrol (DES), an estrogen analogue known to increase the risk of breast cancer in women exposed in utero, is hypothesized to induce neoplasia through a similar genotoxic mechanism. Cultured MCF-10F human breast epithelial cells were treated with DES at varying concentrations and for various times to determine whether the addition of DES to MCF-10F cells resulted in the formation of depurinating adducts. This is the first demonstration of the formation of DES–DNA adducts in human breast cells. A dose-dependent increase in DES–DNA adducts was observed. Demonstrating that treatment of MCF-10F cells with DES, a known human carcinogen, yields depurinating adducts provides further support for the involvement of these adducts in the induction of breast neoplasia.Previous studies have demonstrated the ability of antioxidants such as resveratrol to prevent the formation of estrogen–DNA adducts, thus preventing a key carcinogenic event. In this study, when MCF-10F cells were treated with a combination of resveratrol and DES, a dose-dependent reduction in the level of DES–DNA adducts was also observed.

► The human carcinogen diethylstilbestrol (DES) is metabolized to a reactive DES-quinone.
► DES-quinone reacts with DNA to form 3′-OH-DES-6′-N3Ade and 3′-OH-DES-6′-N7Gua adducts.
► The DES–DNA adducts are formed in MCF-10F human breast epithelial cells cultured with DES.
► The antioxidant resveratrol inhibits formation of DES–DNA adducts in MCF-10F cells cultured with DES.