Reappraisal of eight representative carcinogenic and non-carcinogenic compounds in a new medium-term rat liver bioassay using d-galactosamine

The carcinogenic potential of eight different compounds was assayed in a new medium-term carcinogenicity bioassay using d-galactosamine (DGA) as a non-surgical method to induce regeneration in place of partial hepatectomy (PH). Male rats were initially given a single i.p. injection (200 mg/kg) of diethylnitrosamine (DEN) and after 2 weeks on basal diet, received two i.p. injections of DGA (300 mg/kg) at the end of weeks 2 and 5. They were treated with one of the test compounds aflatoxin B1, benzo[a]pyrene, diethylstilbestrol, urethane, sodium saccharin, bucetin, D-mannitol and sodium chloride in the diet or basal diet alone for weeks 3–8. Carcinogenic potential was assessed by comparing the numbers and areas per cm2 of glutathione S-transferase placental form-positive (GST-P+) foci in the livers of treated animals with those of the control animals given DEN/DGA alone. Positive estimations of carcinogenicity were obtained for the hepatocarcinogens aflatoxin B1, diethylstilbestrols or urethane, and for the non-liver carcinogen benzo[a]pyrene. Negative values were shown in rats treated with the non-carcinogens, d-mannitol and sodium chloride. The two other non-liver carcinogens sodium saccharin and bucetin, also did not exert positive effects in the system. The present data are consistent with findings in previous medium-term bioassays using PH. Our results thus confirm that the present bioassay protocol with repeated administration of DGA instead of PH may offer a new and sensitive non-invasive method to screen large numbers of environmental carcinogens.