1 × 105 HCT116 and CaCo-2 cells were suspended in serum-free medium, then inoculated into the top chamber of transwell chamber (Corning Inc., Corning, NY, USA) pre-coated with matrigel (Corning Inc.). The bottom chamber was added with DMEM medium containing 10% FBS. After 24 h, cells passed into the bottom chamber were fastened in 4% paraformaldehyde fix solution (Beyotime) and dyed using Crystal Violet Staining Solution (Beyotime). The pictures of invasive cells were acquired by an inverted microscope (× 100 magnification; Olympus, Tokyo, Japan) and cell number was counted in the arbitrary three fields.

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