In a first set of experiments, we evaluated the impact of several small molecule inhibitors targeting major signaling hubs of the PI3K/AKT/mTOR and the Ras/RAF/MAPK pathway in DLD-1 colon cancer cells in conventional 2D (3000 cells/well) and 3D spheroid (3000 cells per spheroid) cultures. This evidence concerns the gene PIK3CA and malignant colon neoplasm.