Fig. 5
CR-1 promotes ccRCC cell migration, invasion and angiogenesis in vitro and in vivo. a and b Ectopic expression of CR-1 stimulates cell migration and invasion in Caki-2 cells whereas knockdown of CR-1 inhibits cell migration and invasion in 786-O and Caki-1 cells as determined by Transwell migration (a) and Matrigel invasion assays (b). Data represent the mean ± SD of three independent experiments. **P < 0.01 by two-sided unpaired Student’s t-test. c Ectopic expression of CR-1 enhances cell migration whereas knockdown of CR-1 inhibits cell migration as determined by the wound healing assay. Photomicrographs were obtained at 0 and 24 h and migrated distance was measured using Image J software (Bethesda, USA). Data represent the mean ± SD of three independent experiments. **P < 0.01 by two-sided unpaired Student’s t-test. d Tube formation assay was performed with HUVECs cells incubated in culture supernatant fluid of indicated cell lines in the presence or absence of anti-VEGF neutralizing antibody. Data represent the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01 by two-sided unpaired Student’s t-test. e The new blood vessel formation in CAM was quantified. Data represent the mean ± SD of three independent experiments. **P < 0.01 by two-sided unpaired Student’s t-test. f Relative protein expression of CR-1 and VEGF as determined by Western blot for indicated cell lines. g IHC staining for CD31 and VEGF in tumor specimens from xenografts. Original magnification, × 100. h In vivo metastasis assay was performed in nude mice (5 mice /group) injected with Caki-1/LV-shNC or Caki-1/LV-shCR-1 cells (2 × 106). Left panel, representative images and HE staining of lungs. Original magnification, × 200. Right panel, numbers of tumor nodules in each group. Data represent mean ± SD. **P < 0.01 by two-sided unpaired Student’s t-test