ZA inhibits Edu-macrophages and tumor progression induced by Edu macrophages in the orthotopic XPAI pancreatic cancer mouse model.

<p>(<b>A</b>) Representative fluorescence images of Edu-macrophages after ZA treatment. Scale bars: 10 µm. (<b>B</b>) Bar graphs of the number of Naïve- or Edu-macrophages after ZA treatment in vitro. The numbers of both Naïve- and Edu-macrophages treated with ZA were significantly reduced at every dose. ZA killed both Naïve and Edu in a dose-dependent manner. ** <i>P</i><0.01 (vs. control group). (<b>C</b>) Intravital imaging of XPA1-RFP tumor-bearing mice at the termination of the experiment. Scale bars: 10 mm. (<b>D</b>) The primary tumor weight of Edu-macrophage-treated mice were significantly increased compared to control or Naïve-macrophage-treated mice (control: <i>P</i> = 0.026; Naïve: <i>P</i> = 0.03, respectively). The primary tumor weight of Edu-macrophage + ZA-treated mice were significantly decreased compared to Edu-macrophage-treated mice (<i>P</i> = 0.006). * <i>P</i><0.05, ** <i>P</i><0.01. (<b>E</b>) The metastasis weight of Edu-macrophage-treated mice were significantly increased compared to control or Naïve-macrophage-treated mice (control; <i>P</i> = 0.012, Naïve; <i>P</i> = 0.015, respectively). No metastasis was detected in Edu + ZA-treated mice. There was a significant difference between Edu and Edu + ZA-treated mice (<i>P</i> = 0.025). * <i>P</i><0.05.</p

Imaging of the interactions between host macrophages and cancer cells in live mice.

<p>(<b>A</b>) Scheme of imaging the interactions between host macrophages and cancer cells in live mice. GFP nude mice with dual-color XPA1 subcutaneous tumors were the source of Edu-macrophages. GFP nude mice without tumors were the source of Naïve-macrophage. A skin-flap was spread and fixed on a flat stand. XPA1-GFP-RFP cells (1×10⁶ in 100 µl medium) were sprinkled over the surface of the skin–flap of mice. Twenty-four hours later, the inside surface of the skin-flap was directly observed with the FV1000 confocal microscope (Olympus). Scale bars: 10 mm. GFP host macrophages and dual-color XPA1 cancer cells were observed in Naïve-macrophage mice (<b>B</b>) and Edu-macrophage mice (<b>C</b>). Phagocytosed cancer cells (white arrowheads) and mitotic cancer cells (yellow arrowheads) were detected in both groups. The mean values of phagocytosized and mitotic cancer cells were calculated from four fields with a 20× magnification objective (<b>D and E</b>). Mitosis significantly increased in cancer cells in mice with Edu-macrophages compared to cancer cells in mice with Naïve-macrophages (<i>P</i> = 0.001). More phagocytosis of cancer cells tended to be detected in mice with Naïve-macrophages, (<i>P</i> = 0.061). Scale bars: 20 µm.</p