Dehydroepiandrosterone (DHEA), a hormone of the adrenal cortex, acts as a peroxisome proliferator and hepatocarcinogen in rats upon long-term treatment with high doses in the diet. The aim of the present study was to identify the site of origin of hepatocellular neoplasms and the sequence of preneoplastic lesions. Twenty-five female and 25 male rats were given 0.6% DHEA in the diet; 25 animals of each sex were controls. Groups of 5 treated and untreated animals were sacrificed after 4, 20, 32, 70, and 84 wk. Amphophilic cell foci were detected after 32 wk of treatment; they developed from the liver parenchyma almost exclusively in the vicinity of portal tracts. Adenomas of the amphophilic or amphophilic/tigroid cell phenotype were observed at 70 wk of treatment. Highly differentiated hepatocellular carcinomas presenting a similar cellular phenotype occurred after 70-84 wk. The incidence of hepatocellular carcinomas was 44% in female and 11% in male rats. Ultrastructural studies of the amphophilic cell foci and tumors revealed a marked proliferation of mitochondria and a moderate proliferation of peroxisomes in all lesions. In addition, a very strong peroxisome proliferation was observed in perivenular hepatocytes in the liver of female rats. Peroxisomes usually lacked core and showed flocculent matrices. In male rats, weak peroxisomal proliferation was observed. Typical morphological abnormalities of these peroxisomes were paracrystalline inclusions of striated appearance. Although the most prominent peroxisome proliferation was observed in perivenular hepatocytes, these cells did not seem to be involved in tumor development. In contrast, the morphological similarity of the amphophilic cell foci and the amphophilic/tigroid cell adenomas and carcinomas, their coincident localization near portal tracts, and the sequential appearance of these lesions suggest that the amphophilic cell foci represent an early stage in DHEA-induced hepatocellular neoplasia. Mitochondrial proliferation as the most prominent feature in all stages of this model of hepatocarcinogenesis may offer a new approach for analysis of hepatocarcinogenesis induced by DHEA and possibly other peroxisomal proliferators.

Preneoplastic liver foci and neoplasms of different morphological phenotypes were induced in rats with N‐nitrosomorpholine (NNM; 120 mg/l in drinking water for 7 weeks) and the peroxisome proliferator dehydroepiandrosterone (DHEA; 0.6% in the diet for up to 84 weeks). Preneoplastic glycogen storage foci (GSF) occurred mainly upon treatment with NNM, and amphophilic cell foci (APF) were mainly observed in rats treated with DHEA alone or in combination with NNM. The 2 types of lesions belong to 2 different cellular lineages, the glycogenotic/basophilic lineage and the amphophilic lineage, which are characterized by distinct patterns of alterations in key enzymes of energy metabolism. Whereas in GSF enzymes of glucose metabolizing pathways were modified (increase in glucose‐6‐phosphate dehydrogenase and pyruvate kinase, decrease in glucose‐6‐phosphatase), APF mainly demonstrated alterations in mitochondrial enzymes (increase in cytochrome c oxidase, succinate dehydrogenase and glycerol‐3‐phosphate dehydrogenase) and, to a lower extent, in peroxisomal enzymes (increase in peroxisomal hydratase and acyl‐CoA oxidase). The alterations in enzyme expression reflect an insulinomimetic effect in GSF and a thyromimetic effect in APF. Neoplasms resulting from APF show a more differentiated phenotype than those arising from GSF. We suggest that the different and in many aspects opposite effects of the 2 carcinogens on key enzymes of distinct pathways of energy metabolism modulate the process of neoplastic liver cell transformation and result in phenotypically different preneoplasias and neoplasias reflecting different cellular lineages. Int. J. Cancer (Pred. Oncol.): 79:232–240, 1998.© 1998 Wiley‐Liss, Inc.

Metallic chambers were implanted into the proximal tibiae of rabbits to permit microscopic examination of living bone in situ. The bone repair process secondary to the injury produced during installation of the chamber, was visualized. Six to 8 weeks after implantation, osteoid and/or bone could be seen. The effects of various doses of disodium ethane-1-hydroxy-1, 1-diphosphonate (EHDP) on the repair and regeneration processes following chamber implantation were studied. Data from various techniques indicated that: (1) following low dose EHDP (0.25 mg/kg/day) chambers contained bone tissue morphologically and ultrastructurally indistinguishable from controls; and (2) with higher doses of EHDP (2.5 or 10 mg/kg/day) chamber contained spicules of normal osteoid, osteoblasts and osteocytes, but were devoid of osteoclasts. The effects of the various regimes of EHDP also were assessed on regenerated, trabecular bone contained within the tibia chambers three months after implantation of the chambers. Data from various methods of analysis supported the following conclusions: (1) the low dose of EHDP (0.25 mg/kg/day) had no toxic effects on the trabecular bone within the chambers but there appeared to be an increase in bone formation as compared to saline control; (2) higher doses of EHDP (2.5 or 10mg/kg/day) were not toxic to bone cells but thick osteoid seams formed on the trabecular bone within the chambers. No osteoclasts were found associated with the bone apparently due to the coverage of bone surfaces by osteoid seams; and (3) osteoid which accumulated after EHDP treatment of 2.5 mg/kg/day for 2 months remained uncalcified for as long as 2 months following withdrawal of EHDP administration. The results showed the value of tibial chamber for examining microscopically living bone in situ and demonstrated the inhibitory effect of EHDP on mineralization of newly formed osteoid and a lack of effect on bone cells.