Quantifying Cell Migration and Invasion

Cell migration was determined using a 24-well transwell chamber (Corning, New York, USA), and cell invasion using a 24-well transwell chamber (Corning, New York, USA) coated with 50mg/L Matrigel (BD Biosciences). Briefly, indicated-mimics transfected HeLa and SiHa cells in 100 μL serum-free RPMI 1640 medium (Gibco by Life Technologies, USA) were plated in the top chamber, with a final density of 2×10⁵ cell/well. Next, 600 μL complete medium was added to the lower chambers. Twenty-four hours later, the invaded or migrated cells were fixed with methanol, stained with crystal violet.22 (link) The images in five random fields were taken was by an Olympus BX53 microscope carrying DP72 camera (Olympus, Japan) and cell number was counted by Image J software (NIH). Parallel experiments were carried out five times independently.