Application of bioluminescence imaging of renilla luciferase labelling liver cancer cells in mice in vivo liver cancer tracking

Objective To investigate the application value of bioluminescence imaging of ranilla luciferase (Rluc) labelling liver cancer cells in mice in vivo liver cancer tracking. Methods Rluc-GFP-Hepa1-6 liver cancer cells were constructed. The stable cells with positive expression of GFP were purified by using flow cytometry. Rluc-GFP-Hepa1-6 cells were used for cell bioluminescence imaging. The correlation between the bioluminescence signal intensity and cell quantity was analyzed. The mice model with grafted tumors was constructed by subcutaneous inoculation of Rluc-GFP-Hepa1-6 cells, and in vivo bioluminescent imaging was applied. The growth of tumor cells in the mice was quantitatively monitored. The correlation between the bioluminescence signal intensity, cell quantity and tumor size was analyzed using linear regression analysis. Results The Rluc-GFP-Hepa1-6 cell lines were successfully constructed. The GFP positive rate of Rluc-GFP-Hepa1-6 cells purified by flow cytometry was 95%. Cell bioluminescence imaging revealed that the bioluminescence signal intensity of Rluc-GFP-Hepa1-6 cells was significantly correlated with cell quantity (R2=0.999). In vivo bioluminescence imaging revealed that the bioluminescence signal intensity of subcutaneous tumors was significantly correlated with tumor size (R2=0.887). Conclusions Rluc bioluminescence imaging can successfully monitor the evolution of subcutaneous malignant tumors in mice model with liver cancer. It not only serves as an ideal model for studying the growth, metastasis and treatment of liver cancer, but also provides an excellent, minimally invasive quantitative tracking approach for evaluating the therapeutic efficacy of liver cancer. Key words: Carcinoma, hepatocellular; Fluorescent antibody technique; Bioluminescent imaging; Renilla luciferase