Abstract: OBJECTIVE To establish copepod super green fluorescent protein(copGFP) and nano luciferase(Nluc) labeled MHCC97-H human hepatoma cells. To construct a model of brain metastasis in nude mice, in which can be observed by in vivo imaging system. METHODS The lentiviral vector plasmid carrying the target gene was used to transfect 293FT cells by calcium phosphate method, and the lentivirus was packaged to infect MHCC97-H human hepatoma cells. The positive cells were screened by puromycin, and the monoclonal cell strain stably expressing copGFP and Nluc were purified by limiting dilution assay. The brain metastasis model of nude mice was established by modified carotid artery injection, and the growth of intracranial metastasis was monitored by in vivo imaging system. RESULTS Successfully established a stable strain of MHCC97-H human hepatoma cells which were labeled by copGFP and Nluc. The stable strain and the original strain were used to establish the model by modified carotid artery injection method, and there was no statistically significant difference in body mass change and survival curve between the two groups of tumor-bearing nude mice. The formation of brain metastasis in nude mice with stable strain could be monitored in real time by in vivo imaging system, and the intensity of bioluminescence signal was gradually enhanced with the growth of tumor. CONCLUSION MHCC97-H human hepatoma cells labeled by copGFP and Nluc are successfully established and triumphally constructed a model of brain metastasis in nude mice by a modified carotid artery injection method. This experimental model can be non-invasively and continuously visually monitored by in vivo imaging system, which provides an ideal murine model for the study of the brain metastasis mechanism of liver cancer and the development of therapeutic drugs.