EFFECTS OF 2,3,7,8-TETRACHLORO-P-DIOXIN AND 1,2,3,4,6,7,8-HEPTACHLORODIBENZO-P-DIOXIN ON THE PROLIFERATION OF PRENEOPLASTIC LIVER-CELLS IN THE RAT

Using an initiation - promotion system, enzyme-altered putative preneoplastic liver foci were induced in female Wistar rats by application of diethylnitrosamine (10 mg/kg/day) for 5 days, followed by bi-weekly treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; corresponding to 100 ng/kg/day) or 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HCDD; corresponding to 5 mu g/kg/day) for up to 17 weeks. Groups of animals were killed at various time intervals after start of promoter treatment. For evaluation of DNA synthesis, 5-bromo-2'-deoxyuridine was administered 24 h prior to killing the animals. Quantitative analysis of the number and volume fraction of adenosine-triphosphatase-deficient liver foci revealed that the promoting activity of both dioxins was roughly comparable under the experimental conditions employed. Nuclear labelling indices (LIs) of normal hepatocytes were not altered by TCDD or HCDD treatment, while a slight increase in LIs of non-parenchymal liver cells was observed. Using an immunohistochemical double-staining technique, hepatocytes within glutathione-transferase P-positive liver foci were found to show an approximately 5-to 10-fold higher LI than normal hepatocytes throughout all periods of investigation. During the time course of the experiment, LIs of foci from all treatment groups decreased with time. However, in TCDD-treated rats, and less pronounced in HCDD-treated rats, the initially high rate of proliferation persisted for a greater length of time than in non-dioxin-treated control animals. Assignment of liver foci into four transection size classes revealed that LIs in larger size classes varied considerably, indicating heterogeneity ire the growth behaviour of individual liver lesions. Overall, both dioxins had no effects on the proliferation of normal hepatocytes, while LIs of enzyme-altered liver lesions were slightly enhanced by treatment with TCDD or HCDD. Whether the selective, albeit moderate increase in the proliferation of enzyme-altered liver cells is sufficient to explain the promoting activity of dioxins, or if additional factors (e.g. decrease in death rates of foci cells) are equally important, remains to be determined in further experiments.