Okadaic acid treatment induces DNA adduct formation in BHK21 C13 fibroblasts and HESV keratinocytes.

Okadaic acid (OA), a toxin involved in diarrhetic shellfish poisoning (DSP), has been shown to be a potent tumor promoter in mouse skin and glandular stomach. However, more recent studies tended to show that OA can also act as a genotoxic. In this study, using the 32P-postlabelling method, DNA adduct formation was obtained in two cell lines (BHK21 C13 fibroblasts and HESV keratinocytes) after treatment by OA for 24 h with a dose range between 0.01 and 5 nM. Nineteen adducts were observed with BHK21 C13 cells and 15 with HESV ones. Low doses did not show adduct formation. Intermediate doses have given the most important number of adducts and with higher doses, the number of adducts decreased dose dependently. Ten adducts were similar in the two strains while 9 were specific of BHK21 C13 cell line and 5 of HESV one. The highest total DNA adduct level from origin parts was estimated at 95.6 adducts/10(9) nucleotides for BHK21 C13 fibroblasts (1 nM OA treatment) and 31.1 adducts/10(9) nucleotides for HESV keratinocytes (0.5 nM OA treatment). In this case, the major adduct (number 3) represented 20% for the fibroblastic cell line and 30% for the keratinocytic strain. The genotoxic effect of OA showed in this study should lead to a more careful survey of DSP outbreaks.