The DEN and CCl4 -Induced Mouse Model of Fibrosis and Inflammation-Associated Hepatocellular Carcinoma

Abstract

Human hepatocellular carcinoma (HCC) develops most often as a complication of fibrosis or cirrhosis. While most human studies of HCC provide crucial insights into the molecular signatures of HCC, seldom do they address the etiology of HCC. Mouse models are essential tools for investigating the pathogenesis of HCC; however, the overwhelming majority of cancer models in rodents do not feature liver fibrosis. Detailed in this unit is a protocol for an experimental mouse model of HCC that arises in association with advanced liver fibrosis. The disease model is induced by a single injection of N-nitrosodiethylamine (DEN) followed by repeated administration of carbon tetrachloride (CCl4 ). A dramatic potentiation of liver tumor incidence is observed following administration of DEN and CCl4 , with 100% of mice developing liver tumors at 5 months of age. This model can be employed for studying the molecular mechanisms of fibrogenesis and HCC development, and in cancer hazard/chemotherapy testing of drug candidates.

Keywords: cancer; fibrosis; genotoxic; liver; mechanisms.

Fig. 1. Experimental design, gross pathology, and histopathology of the liver tumors

(A) A single injection of a mutagenic carcinogenic agent N-nitrosodiethylamine (DEN) is followed by repeat dosing with carbon tetrachloride (CCl₄) for up to 14 consecutive weeks. For more detail, see the BASIC PROTOCOL section. (B) Representative photographs of the livers from the animals in each dosing group. (C) Representative photographs of the histopathology of the livers including fibrosis, hepatocellular adenoma and carcinoma.