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

Takeki Uehara1, Igor P. Pogribny2, Ivan Rusyn3

1 Strategic Development Department, Shionogi & Co, Osaka, 2 National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas, 3 Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, North Carolina
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 14.30
DOI:  10.1002/0471141755.ph1430s66
Online Posting Date:  September, 2014
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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, with100% 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. Curr. Protoc. Pharmacol. 66:14.30.1‐14.30.10. © 2014 by John Wiley & Sons, Inc.

Keywords: liver; fibrosis; cancer; mechanisms; genotoxic

     
 
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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1:

  Materials
  • Female pregnant B6C3F1/J mice (Jackson Laboratory) ∼1 week before delivery
  • Diethylnitrosamine (see recipe)
  • Sterile phosphate‐buffered saline (PBS; vehicle)
  • Carbon tetrachloride (CCl 4; see recipe)
  • Olive oil (vehicle)
  • 5‐Bromo‐2′‐deoxyuridine (BrDU; see recipe)
  • Nembutal (Oak Pharmaceutical) for anesthesia or other approved anesthetic
  • Neutral buffered formalin (10%)
  • Paraffin
  • Hematoxylin and eosin
  • Animal scale
  • Disposable plastic syringes (1.0 ml) with needles for i.p. injection
  • Sharp dissecting scissors
  • Heparin‐containing serum gel Z/1.1‐ml centrifuge tubes (Sardstedt)
  • 0.9% NaCl‐moistened filter paper
  • Razor blades
  • Tissue processing/embedding cassettes (any commercial supplier)
  • Screw‐lid containers
  • Light microscope
  • Additional reagents and equipment for the Masson's trichrome procedure (Masson, )
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Figures

Videos

Literature Cited

Literature Cited
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