Wound Healing Assay for Cancer Cells

HCT116 and HT29 cells in the logarithmic phase were used after trypsin digestion and inoculated into a 6-well plate at a density of 6 × 10⁵ cells per well of 2 ml. The cell fusion rate was close to 100% after 24 h of incubation. A 20-μl pipette tip was used to make vertical scratches on the bottom of the 6-well plate (3 strips per well). Then, the plate was washed three times with PBS solution. Following that, a serum-freedrug-containing medium was added for continued incubation. Finally, images were collected at the appropriate time using a Nikon TS2-S-SM inverted microscope and then processed using ImageJ software (version 8.0, Madison, WI, USA) to calculate the proportion of cell-free area in the treated group versus the control groups [16 (link), 17 (link)].