Transwell chambers 8 μm

Dulbecco’s modified Eagle medium was purchased from Invitrogen (CA, USA). Curcumin was obtained from Sigma (c7727, St. Louis, MO, USA); foetal bovine serum (FBS) was purchased from Gibco (CA, USA). Overexpression or inhibition of miR-22 was performed through the transfection of miR-22 mimics or miR-22 inhibitors from RiboBio (Guangzhou, China), and X-treme GENE siRNA was purchased from Roche (Basel, Switzerland). Cell proliferation assays were performed using the EdU Kit obtained from RiboBio (Guangzhou, China) and the MTS Kit purchased from Promega (WI, USA). Transwell chambers (8 μm) were purchased from Corning (NY, USA).

SW1990 cells were plated in 6-well plates (500 cells/well), followed by incubation for 2-3 weeks. Cells were fixed with 4% formaldehyde and stained with crystal violet. A number of colonies with more than 10 cells were counted under the microscope with colony formation rate = (colony number/seeded cell number) × 100% [37 (link)].
Transwell chambers (8 μm, Corning, NY) were applied for cell migration evaluation [38 (link)]. SW1990 cells resuspended with 200 μl serum-free medium were plated in the upper chamber, with 10% FBS medium added in lower chamber. After 24 h, cells were fixed in methanol (Guangzhou Chemical Reagent Factory, Guangzhou, China) for 30 min. The crystal violet- (0.1%) stained cells were counted under an inverted microscope (Caikon Optical Instrument Co. Ltd., Shanghai, China).