Objective To construct a Luc-GFP-labeled human hepatocellular carcinoma ( HCC) cell line with high metastatic potential. In addition, to establish a spontaneous metastasis and conveniently monitored orthotopic model of hepatocellular carcinoma through a cell suspension injection method or a tumor fragment implantation method in nude mice. Methods The recombinant lentiviral expression vector pCDH-CMV-Luc-EF1-GFP-Puro was constructed and transfected into the human hepatoma cell line HCCLM3. The orthotopic xenograft model was established through the cell suspension injection method and the tumor fragment implantation method. The growth and metastasis of the tumors were observed by in vivo imaging and pathological analysis. Results HCCLM3-Luc-GFP, a highly metastatic HCC cell line with GFP expression and Luc activity, was obtained. The tumorigenic rates of the two method were 100%. However, the former can simulate the microenvironment of liver cancer patients, with host factors having less influence. The tumor size and location of the latter are easier to control, with the lung metastatic rate being lower in the former than in the latter. Conclusions The Luc-GFP-labeled highly metastatic human hepatoma cell line HCCLM3-Luc-GFP was successfully constructed. The orthotopic model of highly metastatic and Luc-GFP-labeled HCC in nude mice was successfully established by the above method. This study provides a continuous and effective means to monitor the growth and metastasis of tumors in vivo, for evaluating the efficacy of anti-metastatic drugs against HCC.