title: Perturbation of hepatocyte nuclear populations induced by iron and polychlorinated biphenyls in C57BL/10ScSn mice during carcinogenesis.
abstract: The induction of hepatocarcinogenesis by polychlorinated biphenyls (PCBs) in C57BL/10ScSn mice is markedly potentiated by iron. To investigate the effects of iron and PCBs on nuclear populations, C57BL/10ScSn mice received a single dose of iron-dextran (600 mg Fe/kg) and were fed a diet containing 0.01% of the PCBs mixture Aroclor 1254 for up to 6 months. DNA content of isolated nuclei and hepatocytes was estimated by flow cytometry. Cell suspensions and nuclei isolated from Aroclor treated mice after 6 months contained increased diploid (2N) populations compared to controls. In contrast, iron treatment of mice markedly enhanced fractions of octoploid (8N) nuclei by 2 weeks and this effect persisted over the 6 month period. When Aroclor 1254 and iron were administered together there was a synergistic increase in the mononucleated diploid fraction which was significant at 2 weeks and highly significant at 6 months. This became the predominant nuclear effect. At six months, Aroclor 1254 and iron, both alone and in combination, also increased the rate of DNA synthesis in hepatocytes as measured by bromodeoxyuridine (BrdU) incorporation. The chronic polyploidizing effect of iron overload alone was investigated further and shown to be proportional to the dose and was detectable as early as 2 days after 600 mg Fe/kg and 1 week after 150 mg Fe/kg. Polyploidization of nuclei was inhibited by the oral iron chelator CP94. Iron also induced a prolonged reduction in the incidence of binucleated cells. Histologically, nuclear enlargement due to iron was confined to the midzonal region of the liver lobule, whereas iron deposition was greatest in the periportal region. Iron (600 mg/kg) also caused increased nuclear polyploid states in hepatocytes of adult rats and gerbils. Similarly, weanling mice with a dominantly diploid cell population, when treated with iron (300 mg/kg), exhibited a significant shift to a tetraploid (4N) population and a marked increase in proliferation as measured by BrdU incorporation and proliferative cell nuclear antigen (PCNA) detection. These results indicate that Aroclor 1254 and iron induce changes in the mouse hepatocyte population that involve 2N and 8N nuclei respectively. The combination treatment leads to the emergence and proliferation of a mononucleated, diploid population as observed frequently in chemical hepato-carcinogenesis. The reason for the chronic polyploidizing effect of iron is unknown, but may imply both increased DNA synthesis and impairment of nuclear division with implications in human conditions of iron overload.
authors.affiliation: MRC Toxicology Unit, University of Leicester, UK.
authors.forename: S
authors.initials: S
authors.lastname: Madra
authors.forename: J
authors.initials: J
authors.lastname: Styles
authors.forename: A G
authors.initials: AG
authors.lastname: Smith
mesh_terms: D000818:Animals; D001140:Aroclors / Q000633:toxicity*; D002273:Carcinogens / Q000633:toxicity*; D002467:Cell Nucleus / Q000187:drug effects; D020111:Chlorodiphenyl (54% Chlorine); D004247:DNA / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D005260:Female; D005849:Gerbillinae; D007501:Iron / Q000008:administration & dosage / Q000633:toxicity*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure*; D008114:Liver Neoplasms, Experimental / Q000139:chemically induced* / Q000378:metabolism / Q000648:ultrastructure*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D011003:Ploidies; D011728:Pyridones / Q000494:pharmacology; D051381:Rats; D011916:Rats, Inbred F344; D013045:Species Specificity
chemical_list: D001140:Aroclors; D002273:Carcinogens; D011728:Pyridones; D020111:Chlorodiphenyl (54% Chlorine); C068543:1,2-diethyl-3-hydroxypyridin-4-one; D004247:DNA; D007501:Iron