Cellular kinetics of rat mammary gland terminal end bud epithelium exposed to N-methyl-N-nitrosourea in vivo.

Administration of the direct acting carcinogen N-methyl-N-nitrosourea (NMU) to 50-55-day-old virgin female rats on different days of the estrous cycle yields differential breast tumor biology (T. A. Ratko and C. W. Beattie, Cancer Res., 45: 3042-3047). One basis for these estrous cycle-dependent differences may be the duration of cell cycle stages of susceptible structures such as mammary terminal end buds or the quantity and duration of repair effected following adduct formation within these structures. The terminal end bud (TEB) epithelial cell cycle was characterized using pulse injections of [3H]thymidine (0.5 mCi/g body weight). On estrus, TEB epithelial cell cycle was significantly shorter (15.5 h) than on proestrus (19.9 h) and diestrus (18.8 h). The shorter duration in TEB cell cycle on estrus was likely due to a shorter TG1 (3-4 h) (P less than 0.05) since TS and TG2 did not differ between estrous cycle days. When NMU was injected 1 h after [3H]thymidine, the labeled mitotic wave within TEB of diestrus rats recovered approximately 2-3 h sooner than those given injections during proestrus (P less than 0.01), suggesting less initial damage or a slightly faster rate of DNA adduct repair. When [3H]thymidine was injected 1-5 days after the NMU, the percentage of labeled mitoses of rats given injections during diestrus and proestrus recovered to near normal 48 h after NMU, although the proportion of all cells labeled was still low compared to non-NMU-treated rats. The percentage of labeled mitoses and labeling of cells were normal 3 and 5 days after NMU. Rats receiving a carcinogenic but sublethal dose of NMU (5 mg/100 g body weight), followed by [3H]thymidine injection within 1 min, had one-half the intensity of thymidine incorporation into the terminal end bud DNA of non-NMU-treated rats. Unscheduled DNA synthesis was not demonstrable within the first 48 h following injection of NMU. The results support and extend the finding that rat mammary epithelial cell carcinogenicity of NMU is estrous cycle dependent and appears to be correlated with a differential response in the cell cycle of TEB (shorter at estrus) or delayed recovery in response to NMU (proestrus versus diestrus).