Cell kinetic effects of low doses of the skin carcinogen 7,12-dimethylbenz[a]anthracene on hairless mouse epidermis.

Abstract

Groups of hairless (hr/hr) mice were given a single, topical skin application of either 50, 5 or 0.5 micrograms 7,12-dimethylbenz[a]anthracene (DMBA). At different times up to 3 days after treatment epidermal DNA distribution patterns were determined by flow cytometry, and sp. act. of DNA and labeling indices were obtained based on incorporation of [3H]thymidine. Mitotic rates were determined by the Colcemid method, and the number of basal and suprabasal cells were scored in histological sections. All three doses of DMBA led to an early depression in the uptake of [3H] thymidine, associated with an accumulation of cells with S phase DNA content peaking at 16 h. By combining the methods for studying DNA synthesis, it can be concluded that the alterations observed probably were due to a slow rate of DNA synthesis in the S phase cells, rather than to a block at the entrance of cells into the S phase. Three days after the application, DMBA still maintained an effect on the cell cycle progression, at least in the S phase. There was an obvious dose-response relationship in the inhibition of epidermal DNA synthesis. Six hours after application of the two highest doses of DMBA an early increase in the mitotic rate was observed. This short-lasting high mitotic rate was followed by a transient, very brief increase in the number of suprabasal cells. Thereafter a decrease in both the mitotic rate and the number of suprabasal cells occurred, probably caused by the alterations in DNA synthesis. After the lowest dose there was no such early increase in the mitotic rate and no initial, short-lasting increase in the number of suprabasal cells. Hence, this study shows that decreasing doses of DMBA provoke decreasing degrees of the same type of cell kinetic perturbations in the epidermal cell cycle.