Persistence and accumulation of (potential) single-strand breaks in liver DNA of rats treated with ethyl methanesulphonate.

In vivo alkylation of DNA leads to DNA fragmentation in alkaline sucrose gradients. In a previous paper (Chem.-Biol. Interact., 19 (1977) 111) we presented evidence that, depending on the experimental conditions, a major fraction of the single-stranded breaks observed might be derived from alkali-labile alkylphosphotriesters. Using alkaline gradients the present paper shows that injection of ethyl methanesulphonate (EMS) into Sprague-Dawley female rats results in significantly increased liver DNA fragmentation up to at least 56 days after injection. Accumulation of single-strand breaks was indicated by experiments in which at 6 days after the last of a series of 5 weekly EMS injections (5 X 110 mg/kg) 11.4 breaks/10(9) Dalton were found, being 3 times more than the number of breaks observed at 6 days after a single injection of 110 mg/kg EMS (3.8 breaks/10(9) Dalton). In animals treated with methyl methanesulphonate (MMS) single-strand breaks were observed at 4 h, 1 day and 2 days, but not at 6 days after injection (40 mg/kg). Repeated weekly injections of MMS (5 X 40 mg/kg) did not result in increased numbers of breaks when compared with animals receiving a single injection of this agent (1 X 40 mg/kg; animals were killed 1 day after (the last) injection). It is suggested that MMS-induced breaks are derived, either on the gradient or in situ, from apurinic sites, whereas persistent EMS-induced breaks reflect the presence of ethylphosphotriesters. The results are discussed in relation to the lacking capacity of EMS to induce foci of precancerous lesions in rat liver and the non-hepatocarcinogenic properties of both MMS and EMS.