Objective To compare the tumor growth and metastasis of three different mouse breast tumor cells with Luc-labeled (4T1-Luc, 66c14-Luc and 4TO7-Luc) by using bioluminescent imaging system, and to develop ideal tumor model and living animal analysis technique for tumor metastasis and anti-metastatic therapy. Method The vector containing luciferase gene was constructed and transfected into mouse breast tumor cell line cells and selected with G418 to obtain stable Luc-expressing clones. The cells in logarithmic phase was collected and diluted to 1?07 cells/mL. Then 0.1 mL cell suspension was inoculated into the second mammary fat pad on the right side of normal BALB/c mice to establish the orthotopic models. And another 0.1 mL cell suspension was intravenously inoculated into the tail vein of BALB/c mice to establish the tail vein xenografted model. Their tumorigenesis and metastasis were analyzed in vivo. Result The stable Luc-expressing cell lines were obtained. The whole-body optical imaging found that tumors could be formed after 7 days when the cells were inoculated orthotopically. After 28 days, the tumor sizes of 4T1 were the biggest, the tumor sizes of 66c14 were the second biggest and 4TO7 were the smallest. After 35 days, the tumor sizes were similar among three kinds of cells, while the tumor metastases were showed between 4T1 and 66c14 cells. The metastasis of 4T1 tumor was more serious than 66c14 tumor. 4TO7 tumors didn’t transfer. After 42 days, the tumor sizes were similar among three kinds of cells. The metastasis of 4T1 and 66c14 tumor became more seriously with time passed, and the metastasis of 4T1 tumor was more extensive than 66c14 tumor. 4TO7 tumors still didn’t transfer. After 7 days, the whole-body optical imaging found that tumor could be formed in lung when the stable Luc-expressing cell suspension was intravenously inoculated into the tail vein. The fluorescent signal of 4T1 tumor was the strongest, and deaths of mice occurred in succession. The fluorescent signal of 4TO7 tumor was the second strongest, and 66c14 was the weakest. After 14 days, the metastasis of 4TO7 and 66c14 tumor became more seriously with time passed, and the fluorescent signal of 4TO7 tumor was stronger than 66c14 tumor, and deaths of mice occurred in succession. Conclusions Breast spontaneously metastatic model could show the metastatic characteristics and the whole metastatic process more authentic than the tail vein xenografted model，which is an ideal model for the study of tumor metastasis.