To establish animal models of liver tumors, the PLCε gene knockout mice was induced by mutation inducers Diethylnitrosamine (diethylnitrosamine DEN) combined with tumor enhancer phenobarbital (phenobarbital PB). Methods 12-day-old male PLCε-/ - and PLCε + / + mice were randomly selected as group I (n= 40)and group II (n= 40)，respectively．The mice of group I and II were injected DEN through abdominal cavity at the age of 12 days，then drinking PB after 4 weeks and continuously until 24 weeks. At the same time, 12-day-old male PLCε-/ - and PLCε + / + mice were randomly selected as the control group I (n= 40) and control group II (n= 40), respectively, and fed with normal feeds for 24 weeks. At the end of the experiment, the liver tumor was observed under electron microscopy. Results Pathology results confirm that 18 mice of group II had liver tumors, and rate of tumor formation is 60%; 15 mice of group I had liver tumors, and rate of tumor formation is 46.9%; the mice of control group I and II had not liver tumors. Conclusions The animal model of liver tumor have been successfully established by using mutation inducers DEN combined with PB in PLCε gene knockout mice, and could provide a foundation for further researching effects of PLCε during liver tumor formation.