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A modified initiation-selection procedure for neonatal male and female rat hepatocarcinogenesis were examined utilizing the methods of quantitative stereology. In this study, diethylnitrosamine (10 mg DEN/kg) was given a few days after birth. At weaning, the rats were fed 0.02% 2-acetylaminofluorene (AAF) for 2 weeks with a mitotic stimulus [70% partial hepatectomy (PH)] after 1 week on the diet. Quantitative stereological analyses in conjunction with the use of several enzyme markers were used to determine the number and volume of altered hepatic foci (AHF) detected at 1 week, 3 months and 7 months after the selection procedure. This format resulted in an equivalent number of AHF in male and female rats. The AHF were three times larger in males than in females 1 week after discontinuation of AAF administration. Three months after the selection procedure, the number of AHF had decreased by at least a third and their volume percentage was the same in male and female rats. After 7 months, the number and volume fraction of detectable AHF in females were comparable to those which had been observed at 1 week after selection. In the male, the number but not the volume fraction were similar at 7 months compared with 1 week after selection. Both initiation with DEN and selection with AAF/PH contribute independently to the total population of AHF in male and female rats. At least half of the AHF detected 7 months after the selection protocol were due to DEN administration alone. Rats receiving only the AAF/PH selection exhibited one third of the number of AHF observed with the complete protocol. Administration of a non-necrogenic dose of DEN to neonatal rats when coupled with the AAF/PH selection procedure resulted in a significant promotion of the growth of initiated hepatocytes at 1 week, 3 months or 7 months after the selection procedure. These studies demonstrated that (i) the number of AHF detected after a non-necrogenic dose of DEN during the first week of life with subsequent AAF/PH selection after weaning decreases within the first 3 months after the selection procedure, but can re-develop with a promotion stimulus; (ii) the AAF/PH selection procedure itself may initiate hepatocytes in the absence of DEN administration; (iii) the AAF/PH selection procedure is equally effective with respect to the number of AHF observed after phenobarbital promotion in weaning male and female rats initiated near birth.

Quantitative stereological studies of a 'selection' protocol of hepatocarcinogenesis following initiation in neonatal male and female rats