Cell Proliferation, Migration, and Invasion Assays

4.0×10³ cells were seeded into 96-well-plate in quintuplicate. After 72 hours of incubation, the absorbance was measured at 450 nm by adding a CCK8 reagent (Boster Biological Technology) to detect cell proliferation. Cell migration was evaluated by scratch assays as described previously [25 (link)]. After 16 hours of incubation, the migrated area was photographed and calculated by ImageJ software. For invasion assay, 8 mm pore transwell inserts (Corning Life Sciences) were coated with 75μl Matrigel matrix (Corning, 200 μg/mL) as upper chambers. 700μl DMEM medium with 20% FBS was added into the 24-well-plate. Meanwhile, 5.0×104 cells were suspended in a serum-free medium and were added to the inserts which were placed in a 24-well-plate. After 48h, the cells were fixed with 10% formalin and stained with 0.1% crystal violet solution. The numbers of invaded cells were counted at 200x magnification.