Fig. 4

Genipin suppresses HCC cell invasion and reverses EMT process. (a) MHCC97L and HepG2 cells were cultured in trans-well inserts (upper chamber). Images were obtained via optical microscope after genipin treatment 12 h later; invaded cells were calculated by three individual researchers. Scale bar 50 μm. (b) MHCC97L cells were cultured in FITC-conjugated gelatin (green) for 24 h. Phalloidin was applied to stain F-actin (red) and DAPI used to indicate nuclei (blue). Black area underneath the cell indicates gelatin degradation area; scale bar 20 μm. (c) To construct the 3D culture system, MHCC97L cells were seeded into a layer of Matrigel. Then, different dosages of genipin with DMEM and 15% Matrigel were added. Upper mixture was replaced every 24 h. Arrows indicate tubular structure formation on Matrigel. Images were obtained via optical microscope after genipin treatment 96 h later; scale bar 50 μm. (d) Protein expressions of vimentin, fibronectin, N-cadherin, and E-cadherin were validated by western blot and immunofluorescence staining (green). Nuclei were stained by DAPI (blue). Scale bar 20 μm. Data presented as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001