Our results revealed that transfection with Twist1 cDNA expression plasmid significantly promoted si-AFAP1-AS1-induced inhibition of cell migration, invasion and EMT, as indicated by an increased expression of mesenchymal cell markers (N-cadherin, Vimentin) and a decreased expression of epithelial cell markers (E-cadherin) in osteosarcoma cell lines (Fig. 5a, b and c). This evidence concerns the gene TWIST1 and osteosarcoma.