Figure 8

Human epithelial and mesenchymal liver cancer cells. (A) Representative phase-contrast images (20 × magnification) of Huh7 and MHCC97-L cells. (B) Representative images (4 × magnification) of Huh7 and MHCC97-L wound healing assay after 24 hours of scratching. Experiments were performed in triplicates. (C) Relative expression of mRNA endcoding SNAIL1, ZEB1 and ZEB2 normalized to endogenous control GAPDH. Bars represent mean ± SEM of triplicates, *p < 0.01. Western blot analysis of E-Cadherin and β-actin. Data representative of two independent experiments. (D) Matrigel invasion assay. Phase-contrast images (4 × magnification) of cells that have invaded the membrane and adhered to the bottom of the plate. Data are expressed as the total number of invasive cells at the bottom of the plate, with five fields counted per well, n = 3 wells/cell line, and reported as the mean ± SEM, *p < 0.01. (E) Relative expression of mRNA encoding TISC genes BMI-1, NANOG, OCT4, ABCG2, CD44, CXCR4, and EPCAM normalized to endogenous control GAPDH. Data represent mean ± SEM of triplicates, *p < 0.01. (F) Tumor-sphere assay was performed for two weeks using non-adherent plates. Phase contrast images are representation of three independent experiments (4 × magnification).