Department of Medicine, Division of Gastroenterology; UMass Metabolic Network
Digestive System Diseases | Gastroenterology | Hepatology | Neoplasms
AIM: To establish a mouse model of alcohol-driven hepatocellular carcinoma (HCC) that develops in livers with alcoholic liver disease (ALD).
METHODS: Adult C57BL/6 male mice received multiple doses of chemical carcinogen diethyl nitrosamine (DEN) followed by 7 wk of 4% Lieber-DeCarli diet. Serum alanine aminotransferase (ALT), alpha fetoprotein (AFP) and liver Cyp2e1 were assessed. Expression of F4/80, CD68 for macrophages and Ly6G, MPO, E-selectin for neutrophils was measured. Macrophage polarization was determined by IL-1beta/iNOS (M1) and Arg-1/IL-10/CD163/CD206 (M2) expression. Liver steatosis and fibrosis were measured by oil-red-O and Sirius red staining respectively. HCC development was monitored by magnetic resonance imaging, confirmed by histology. Cellular proliferation was assessed by proliferating cell nuclear antigen (PCNA).
RESULTS: Alcohol-DEN mice showed higher ALTs than pair fed-DEN mice throughout the alcohol feeding without weight gain. Alcohol feeding resulted in increased ALT, liver steatosis and inflammation compared to pair-fed controls. Alcohol-DEN mice had reduced steatosis and increased fibrosis indicating advanced liver disease. Molecular characterization showed highest levels of both neutrophil and macrophage markers in alcohol-DEN livers. Importantly, M2 macrophages were predominantly higher in alcohol-DEN livers. Magnetic resonance imaging revealed increased numbers of intrahepatic cysts and liver histology confirmed the presence of early HCC in alcohol-DEN mice compared to all other groups. This correlated with increased serum alpha-fetoprotein, a marker of HCC, in alcohol-DEN mice. PCNA immunostaining revealed significantly increased hepatocyte proliferation in livers from alcohol-DEN compared to pair fed-DEN or alcohol-fed mice.
CONCLUSION: We describe a new 12-wk HCC model in adult mice that develops in livers with alcoholic hepatitis and defines ALD as co-factor in HCC.
Alpha-fetoprotein, Liver tumor, Macrophage polarization, Proliferating cell nuclear antigen, Steatohepatitis
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DOI of Published Version
World J Gastroenterol. 2016 Apr 28;22(16):4091-108. doi: 10.3748/wjg.v22.i16.4091. Link to article on publisher's site
World journal of gastroenterology
Ambade A, Satishchandran A, Gyongyosi B, Lowe P, Szabo G. (2016). Adult mouse model of early hepatocellular carcinoma promoted by alcoholic liver disease. Open Access Publications by UMMS Authors. https://doi.org/10.3748/wjg.v22.i16.4091. Retrieved from https://escholarship.umassmed.edu/oapubs/2849
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This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License