Fig. 1
RvD1 impedes the CAFs-induced cancer stemness and EMT of HCC cells. (a) Hep3B and SMMC-7721 cells were incubated with CM from CAFs (CMCAFs) or CM from CAFs pre-treated with RvD1(400 nM) (CMCAFs + RvD1) for 24 h, The cells were then seeded in a matrigel-coated invasion chamber for 24 h. The invasive cells were quantified by counting the number of cells in 5 random fields at × 100 magnification. Scale bars = 50 μm. n = three independent experiments, **P < 0.01 by ANOVA. (b) Hep3B and SMMC-7721 cells were treated with CMCAFs and CMCAFs + RvD1 for 48 h, and western blotting analysis was performed to test the expression of stemness markers (OCT4, Nanog, Sox2), and epithelial-mesenchymal transition markers (E-cadherin, N-cadherin, vimentin). (c) Hep3B and SMMC-7721 cells were incubated with CMCAFs and CMCAFs + RvD1 for 24, 48, 72 and 96 h, and cell viability was evaluated by MTT assay. n = three independent experiments, *P < 0.05 or **P < 0.01 by ANOVA. (d) Effects of CMCAFs and CMCAFs + RvD1 on the colony-forming ability of Hep3B and SMMC-7721 cells were assessed by colony formation assay. Images are representative of three independent experiments, and the colony number was counted and plotted. Scale bar = 1 cm. n = three independent experiments, **P < 0.01 by ANOVA. (E) Tumorsphere formation assay of Hep3B and SMMC-7721 cells inculated with CMCAFs or CMCAFs + RvD1. The number and size of tumorspheres were counted, measured and plotted. Magnification is × 200, and scale bars = 50 μm. n = three independent experiments, **P < 0.01 by ANOVA