Building on our previous findings—which demonstrated that LE135, a dual agonist of the mechanosensitive ion channels TRPV1 and TRPA1, could chemically replicate the cellular effects of mechanical stimulation in human osteosarcoma cells [42,48]—the present study sought to further investigate the chemical modulation of MS channel-dependent mechanosignaling in OS by including agonists for both channels, thereby clarifying their individual and combined contributions to this signaling pathway. This evidence concerns the gene TRPV1 and osteosarcoma.