Biliary Proliferation and Adaptation in Furan-Induced Rat Liver Injury and Carcinogenesis

Abstract

Distinctive intrahepatic biliary adaptation responses occur in the liver of rats subjected to select hepatotoxic and/or carcinogenic treatments with the nongenotoxic cholangiocarcinogenic agent furan. Specifically, metaplastic small intestinal-like glands closely resembling in their cellular composition the crypts of Lieberkühn of normal rat small intestine were selectively derived from putative hyperplastic bile ductule-like progenitor structures in the right and caudate liver lobes of young adult Fischer-344 male rats given furan by gavage at a daily dose of 30-45 mg/kg body weight, 5 times weekly, over a 2-6-wk treatment period. Longer term chronic administration of furan at 30 mg/kg/day for 9-19 wk resulted in the preferential development of primary hepatic adenocarcinomas, which arose at 70-100% incidences from right/caudate liver lobes and which were characterized by small intestine mucosal cell differentiation. Interestingly, the neoplastic glands of these "intestinal-type" hepatic tumors demonstrated strongly positive immunochemical reactions for both hepatocyte growth factor/scatter factor and its c-met encoded receptor and were immunohistochemically positive for transforming growth factor β₁ (TGF-β₁) and for mannose-6-phosphate/insulin-like growth factor II receptor, implicated in the activation of latent TGF-β₁. In contrast, a different pattern of aberrant bile ductular cell differentiation was noted to occur in the atrophied right liver lobe of moribund Fischer-344 male rats that were chronically exposed over a 10-14-day period to a severely hepatotoxic dose of furan (60 mg/kg/day). Under this latter experimental condition, rare yet distinct cholangiolar-like structures composed of biliary epithelial cells and typically a single ductular hepatocytic cell in various stages of maturation specifically formed in association with an extensive hyperplastic bile ductular reaction. Very similar cholangiolar-like structures also appeared in areas of preexisting hyperplastic bile ductule tissue at 3-5 days following the administration of a single hepatonecrogenic dose of CCl₄ to rats that 4-6 wk earlier had been subjected to a bile duct ligation. In addition, a novel rat model was developed in which furan combined in a unique synergistic manner with bile duct ligation to induce the replacement of almost all of liver with well-differentiated hyperplastic bile ductules without evidence of differentiation along either metaplastic small intestine mucosal cell or ductular hepatocyte lineages. Bile ductular epithelial cell isolates from bile duct-ligated/furan-treated rats were further observed to be organized in the form of bile duct-like structures in vitro under specific conditions of primary cell culture and in vivo following their cell transplantation into the inguinal fat pads of syngeneic recipient rats. Overall, these findings serve to exemplify the remarkable plasticity that may be exhibited by certain proliferating biliary cell populations in liver in response to specific types of severe hepatic injury and/or during cholangiocarcinogenesis.