Fig. 6

VEGF driven-hemangiogenesis and lymphangiogenesis are involved in anti-metastasis activation of miR-7 in GC. a and b miR-7 expression inhibited hemangiogenesis and lymphangiogenesis in vivo. Experimental metastasis mice from HGC-27 and MKN-28 cells were established and metastatic lung and liver tissues were collected as described in Materials & Methods. Hemangiogenesis was evaluated in metastatic lung and liver by IHC staining for CD34 hemendothelial marker (a). Lymphangiogenesis was evaluated by detecting Lymphatic vessels density through IHC staining for LYVE-1 (b). Representative IHC images are shown. Data are presented as mean ± SD. c-d miR-7 alleviated VEGF expression in vitro and in vivo. VEGF expression in GC cells were detected by flow cytometry (c). IHC staining was used to evaluate VEGF expression in metastatic lung and liver tissues (d). Representative IHC images are shown. e miR-7 alleviated VEGF-A and VEGF-C expression. VEGF-A and VEGF-C mRNA were detected by Real-Time PCR. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 between the indicated two groups determined by paired student’s t test. Scale bars: (main) 50 μm; (inset) 200 μm