Diethylnitrosamine combined with carbon tetrachloride to induce the primary hepatic carcinoma model in mice

Abstract：Objective:To establish primary hepatic carcinoma model in mice by diethylnitrosamine (DEN) combined with carbon tetrachloride (CCl4),thus lay the foundation for further study of hepatocellular carcinoma (HCC).Methods:Sixty C57BL/6J mice were randomly divided into normal group (20) and model group (40).The model group mice were intraperitoneally injected with DEN 2 mg· kg-1 at 14 d after birth and intraperitoneally injected with 20％ carbon tetrachloride (CCl4) solution at 5 mL· kg-1 2 weeks later for twice a week and persisting for 16 weeks.At the 20th week,the mice in each group were sacrificed and liver specimens were removed and weighed.The serum activities of alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (ALP) and alpha fetal protein (AFP) were determined by ultraviolet spectroscopy.Liver tissue sections were examined under Hematoxylin and Eosin (HE) staining.Results:Compared with normal group,the serum ALT,AST,ALP,AFP,nodule number,and liver index of the model group mice increased significantly (P ＜ 0.01).Meantime,33 of 40 mice developed HCC (82.5％) and 3 of them died of toxic hepatitis and acute hepatocellular necrosis.The liver in the normal mice showed no obvious abnormalities.Conclusion:Combination of DEN and CCl4 successfully established primary liver cancer model in mice,which provides a new way for liver cancer research.