In vivo anti-metastasis activity in 4T1 tumor-bearing mice

When the tumor volume of 4T1 tumor-bearing mice grew up to 100-150 mm3, the mice were randomly divided into eight groups (n = 13 per group) and then intravenously injected with PBS, Mn-TCPP, MOF or Mn-MOF at Zr dosage of 5 mg/kg and Mn dosage of 1 mg/kg. At 24 h after injection, the mice were or were not irradiated with US (1.0 MHz, 1 W/cm2, 50% duty cycle) for 5 min. The body weights were measured by using a scale-balance, and tumor lengths and widths were measured with a caliper every day. Tumor volume was calculated by the following equation: V= (tumor length)×(tumor width)2/2. On day 20 after treatment, 5 mice each group were sacrificed. Blood samples were collected for serological analysis. Lung tissues were weighed and the metastatic nodules on lungs were counted. Tumors and other major organs (heart, live, spleen, lung and kidney) were harvested and fixed in a 4% paraformaldehyde for H&E staining. Tumor sections were immunofluorescence stained using Cy3-conjugated anti-CD31 antibody, Cy3-conjugated anti-VEGF antibody and Cy3-conjugated anti-HIF-1α antibody. The rest mice were used for long-term survival observation.

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