Estragole DNA adduct accumulation in human liver HepaRG cells upon repeated in vitro exposure

Accumulation of N²-(trans-isoestragol-3'-yl)-2'-deoxyguanosine (E-3'-N²-dG) DNA adducts derived from the alkenylbenzene estragole upon repeated dose exposure was investigated since the repair of this adduct was previously shown to be inefficient. To this end human HepaRG cells were exposed to repeating cycles of 2 h exposure to 50 μM estragole followed by 22 h repair to mimic daily exposure. The E-3'-N²-dG DNA adduct levels were quantified by LC-MS/MS after each cycle. The results show accumulation of E-3'-N²-dG DNA adducts at a rate of 17.53 adducts/10⁸ nts/cycle. This rate at the dose level calculated by physiologically based kinetic (PBK) modeling to result in 50 μM was converted to a rate expected at average human daily intake of estragole. The predicted time estimated to reach adduct levels reported at the BMD10 of the related alkenylbenzene methyleugenol of 10-100 adducts /10⁸ nts upon average human daily intake of estragole amounted to 8-80 (in rat) or 6-57 years (in human). It is concluded that the persistent nature of the E-3'-N²-dG DNA adducts may contribute to accumulation of substantial levels of DNA adducts upon prolonged dietary exposure.