Evaluating Invasive Potential of Cancer Cells

Invasive potential of 143B-GFP-Luc cells, treated with vehicle, or 12 nM miR-34a, or 40 nM doxorubicin plus 4 μM sorafenib, or the combination for 72 h, was assessed as described previously [16 (link), 17 (link), 45 (link)] by using Corning BioCoat Matrigel Invasion Chamber with 8.0 mm PET membrane coated with matrigel (Corning, NY, USA). Briefly, 5 × 10⁴ cells in 500 μl serum-free RPMI 1640 media were placed into the upper insert, and 750 μl medium supplemented with 10% FBS was added into the well as a chemo-attractant. 30 h later, cells in the upper inserts were removed and the lower were fixed with 10% formalin, stained with 0.1% crystal violet, and then photographed with five fields per insert under an Olympus IX2-UCB microscope (40 × magnifications). Each treatment group was conducted in triplicate and assessed independently. The quantification of invaded cells was calculated using Image J software and invasion capacity was determined by adjusting the vehicle control group to 100%.