Luc-4T1cells (5×105) were inoculated subcutaneously into right legs of female Balb/C mice. When tumor volume reached ~50 mm3, mice were randomly divided into 5 groups (n=6): (1) saline group; (2) Met group; (3) SN38 group; (4) Met/SN38 group; (5) MS NPs group. Drugs were intravenously injected via tail vein once every other day for 12 days. To relief the burden of mice, we removed the primary tumor of mice on the 30th day. The Ami X optical imaging system (Spectral Instruments Imaging Co., USA) was used to image spontaneous metastases of cancer cells on days 8, 24, 40. Before in vivo bioluminescence images, D-luciferin sodium salt (150 mg/kg) was given via intraperitoneal injection. After the last imaging, the lungs and tumor of mice were removed. To examine lung metastasis, we took the picture of ex vivo of lungs and counted the number of lung nodules numbers.

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