Estrogen-nucleic acid adducts: Dissection of the reaction of 3,4-estrone quinone and its radical anion and radical cation with deoxynucleosides and DNA

Previous studies from our laboratory have shown that 3,4-estrone quinone (3,4-EQ) can redox-cycle and is capable of inducing single-strand DNA breaks in MCF-7 breast cancer cells, as well as reacting with various deoxynucleosides to give several estrogen-nucleic acid adducts. While reactions of 3,4-EQ with all the deoxynucleosides under acidic conditions gave only the N7-Gua adduct, which could proceed by Michael addition, reactions of 3,4-EQ under reductive conditions gave several adducts, including the N7-Gua, C8-Ade, C8-Gua, NS-Thy, and N4-Cyt adducts, suggesting the involvement of a 3,4-EQ radical species. The question as to which of the reactive species, the estrogen quinone or the estrogen semiquinone, that is responsible for estrogen's genotoxic activity has been the subject of recent investigations in several laboratories. To explore this in more detail, we carried out studies on the reactivity of 3,4-EQ, the 3,4-EQ radical anion, and the 3,4-EQ radical cation with both deoxynucleosides and calf thymus DNA under different pH conditions. Both stable and unstable adducts with guanine and thymine were observed from reactions with DNA. Although adduct levels were somewhat different, the adduct profiles obtained from reactions of 3,4-EQ and its radical anion with both DNA and deoxynucleosides were quite similar and were found to be significantly different from product profiles obtained from reactions with the 3,4-EQ radical cation. Studies conducted with the human breast tumor cell line MCF-7 demonstrated the formation of the N7- and C8-Gua adducts in which the profiles were similar to those obtained from reactions of 3,4-EQ with DNA. These results suggest that the reactive species that is responsible for adduct formation under physiological conditions is most likely to be the 3,4-EQ radical anion.