Fig. 6.

MiR-25-3p promotes CRC metastasis. a Effects of condition media derived from SW480/mock, SW480/miR-25-3p and HCT116 NC, HCT116/miR-25-3pon permeability of HUVEC monolayers by in vitro permeability assay. Mean ± SEM are provided (n = 3). b Top, effects of condition media derived from SW480/mock and SW480/miR-25-3p on tube formation ability of HUVECs by tube formation assay. Bottom, effects of condition media derived from HCT116 NC and HCT116/miR-25-3p on tube formation ability of HUVECs. Scale bar represents 100 µm. Mean ± SEM are provided (n = 3). c Top, effects of condition media derived from SW480/mock and SW480/miR-25-3p on vascular outgrowth of rat aortic rings. Bottom, effects of condition media derived from HCT116 NC and HCT116/miR-25-3p on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided (n = 3). d Effect of miR-25-3p knockdown on liver metastasis of mice. HCT116 NC or HCT116/miR-25-3p was injected into the mesentery at the tail end of cecum. The number of lung metastatic sites (indicated by arrows) was counted under the microscope. Mean ± SEM are provided (n = 5). Scale bar in left panels represents 1 cm. Scale bar in middle panels represents 0.5 cm. Scale bar in right panels represents 100 µm. e Effect of miR-25-3p knockdown on introtumoral microvessel density in primary and metastatic tumor. The number of microvessel was counted under the microscope. Mean ± SEM are provided (n = 5). Scale bar represents 50 µm. f Effect of miR-25-3p knockdown on hepatic and pulmonary vascular permeability in primary and metastatic tumor. The liver and lung were derived from mice 30 days after being orthotopically implanted with HCT116/zip-miR-25-3p cells. Levels of rhodamine–dextran fluorescence in tissues were quantified using Image J software and normalized to the levels of DAPI. Mean ± SEM are provided (n = 5). Scale bar represents 50 µm. **P < 0.01, ***P < 0.001, ****P < 0.0001 according to two-tailed Student’s t test