Mice exposed prenatally to diethylstilbestrol (DES-exposed mice) can transmit a carcinogenic influence to the next generation (DES-lineage mice) when mated to control mice. The persistence of this effect was studied one generation further (DES-lineage-2 mice) by mating DES-lineage female mice to control males. The interaction of maternal dietary fat levels with DES was also tested by feeding high and low levels of dietary fat during the pregnancies that produced the final two generations. DES-lineage-2 mice, exposed to low or high fat maternal diets, had significantly more tumors than control mice with corresponding dietary fat exposure. The frequency of tumors in DES-lineage-2 mice was not significantly lower than in DES-lineage mice from a previous experiment. Thus, the multigenerational effect of DES is relatively intense in mice. If this type of carcinogenesis can occur in the human population, it poses a major threat to future generations.