Abstract: Objective To establish a human hepatocellular carcinoma cell line with stable and high-level GFP/Luciferase(Luc) expression, and observe the development of tumors in deep organisms of the nude mice by in vivo biofluorescence imaging. Methods The SMMC-7721 -GFP/Luc cell line were established by lentivirus transfection，then were orthotopic implanted in the liver of the BALB/c nude micc, the behaviors of the tumors was detected by in vivo bioluminescence imaging. Results The cell line SMMC-7721-GFP/Luc with stable and high-level GFP/Luc expression had been successfully established. After the cell line had been inoculated in the liver of the nude mice, the tiny tumor masses in early time and the growth of tumors in late time in the deep organisms were observed by in vivo biofluorescence imaging. Conclusion The use of cell line SMMC-7721 -GFP/Luc could directly observe the tumor growth and metastasis in the orthotopic implantation animal model. It would help us to understand the biological behavior of human hepatocellular carcinoma and its characteristics. In view of its sensibility, intuitiveness, dynamics and reliability, it would do some good to further study of human hepatocellular carcinoma.

Key words: GFP/Luc double-tagging, Lentivirus carrier, In vivo bioluminescence imaging, Orthotopic implantation, Human hepatocellular carcinoma