Once the autophagy was blocked by autophagy inhibitors, chloroquine, the cleaved caspase 3 and Annexin V positive cells were reduced (Figure 6A,B), and cell viability was rescued (Figure 6C) in Licochalcone A-treated HOS cells, indicating that the autophagy is associated with Licochalcone A-induced apoptosis in osteosarcoma HOS cells. The gene discussed is ANXA5; the disease is osteosarcoma.