An increased risk of cancer of the esophagus has been reported in alcoholics and in populations with low dietary vitamin A consumption. As cancer is a disorder of cell proliferation and differentiation, we have assessed the combined effects of ethanol and vitamin A deficiency on cell proliferation and structure of the esophagus. Weanling male rats were fed liquid diets with either a standard amount of vitamin A or lacking vitamin A for 8 wk. Littermates were pair-fed the same diets with carbohydrate (36% of calories) replaced by ethanol. Rats were given [3H]thymidine 1 h before death, and the labeling index of the proliferative basal cells was determined on radioautographs. In rats fed the normal vitamin A diet with or without ethanol, plasma vitamin A was normal. Hepatic vitamin A was markedly decreased, whereas esophageal vitamin A was increased after ethanol feeding. Ethanol feeding resulted in a twofold increase in basal cell labeling index (14.6 +/- 0.7 vs. 6.8 +/- 0.8; p less than 0.001). The thickness of the epithelium and the morphology of basal cells were not altered by ethanol feeding. In rats fed the vitamin A-deficient diet with or without ethanol, plasma vitamin A was extremely low, and hepatic and esophageal vitamin A were unmeasurable. The epithelium was thin (with a 50% reduction in thickness) and showed abnormalities of basal cells and increased production of keratohyalin granules, changes suggesting a disorder in the epithelial differentiation. This altered differentiation caused by vitamin A deficiency was not affected by ethanol consumption. Ethanol feeding again resulted in an increase in the basal cell labeling index (13.2 +/- 1.6 vs. 4.8 +/- 0.7; p less than 0.001). Vitamin A deficiency had no effect on basal cell proliferation. Therefore, the stimulatory effect of ethanol on cell proliferation is independent of vitamin A deficiency. Nevertheless, the combined actions of ethanol and vitamin A deficiency may have a synergistic effect on the susceptibility of the esophagus to carcinogens.