Figure 5.

Antitumor immunotherapeutic effect in mice bearing subcutaneous 4T1 grafts. (A) Schematic illustration of immunotherapeutic studies in vivo. (B) In vivo Dir fluorescence imaging of mice bearing the 4T1 tumor after tail vein injection of ST-Dir or NT-Dir and (D) relative Dir fluorescence intensity in tumor tissues at different time points. (C) Ex vivo Dir fluorescence imaging and (E) relative Dir fluorescence intensities in major organs (heart, He; liver, Li; spleen, Sp; lung, Lu; kidney, Ki; tumor, Tu) of the above mice sacrificed at 48 h postinjection. (F) Orthotopic 4T1 tumor growth in mice receiving different treatments (n = 6). The injections were performed every 3 days for a total of seven treatments. (G) Cumulative survival of 4T1 tumor-bearing mice receiving difference treatments (n = 8). (H) Histological analyses of 4T1 tumor sections from mice having undergone a total of 19 days of treatment. In H&E staining, blue indicated nuclei, and red indicated extracellular matrix and cytoplasm. In the TUNEL assay, apoptotic cells were stained brown. Scale bars represent 50 μm. (I) In vivo bioluminescence imaging (BLM) of Luc-4T1 tumor-bearing mice receiving various treatments at different treatment time points to track orthotopic tumor growth and metastases (n = 6). (J) Tumor metastasis potential in mice bearing subcutaneous 4T1 grafts. Representative ex vivo bioluminescent images (BLM) and the H&E staining of livers and lungs obtained from the Luc-4T1 tumor-bearing mice having undergone a total of 19 days of treatment to track tumor metastases. Yellow arrows indicated metastatic tumor nodules. Scale bars represent 50 μm. siRNA dose, 250 μg/kg body weight; AS dose, 60 μg/kg body weight. *P < 0.05; **P < 0.01; ***P < 0.001.