SHARP1 inhibits migration, invasion, and metastasis of EC cells. (A) Overexpression of SHARP1 in Ishikawa and HEC-1B cells revealed by qPCR (left) and western blotting (middle), and blots were further quantified by densitometry of triplicate blots (right), β-actin was included as an internal control. (B) Wound-healing assays for Ishikawa and HEC-1B cells. Representative images were obtained at 100× magnification. Graphs show the relative migration distance after 12 h incubation. (C) Trans-well invasion assays for Ishikawa and HEC-1B cells. Representative images were obtained at 200× magnification. Graph shows the number of invasive cells for each treatment group (averaged across five random images). (D) IshikawaControl or IshikawaSHARP1 cells were injected intravenously into SCID mice. Sections of lung tissue were stained with hematoxylin and eosin. (M) metastatic tumor, (N) normal lung tissue. Images were obtained at 40× (left) magnification. Graph shows the number of visible tumor nodules within analyzed lung tissue (n = 6 per group). Data represent the mean ± SD from one representative experiment of three independent experiments. **P < 0.01, ***P < 0.001.