6-Methoxydihydrosanguinarine induces apoptosis and autophagy in breast cancer MCF-7 cells by accumulating ROS to suppress the PI3K/AKT/mTOR signaling pathway

6-Methoxydihydrosanguinarine (6-MDS) is a natural benzophenanthridine alkaloid extracted from Hylomecon japonica (Thunb.) Prantl. It is the first time to explore the effect and mechanism of 6-MDS in breast Cancer. Network pharmacology, molecular docking, and molecular dynamics simulation technology were adopted to identify the potential targets and pathways of 6-MDS in breast Cancer. Besides, cell proliferation, Apoptosis, and western blotting assays were conducted to investigate the effect of 6-MDS on MCF-7 cells. Network pharmacology, molecular docking, and molecular dynamics simulation results confirmed the effect of 6-MDS on resisting breast Cancer via the PI3K/Akt/mTOR signaling pathway. In addition, the functional experiments results demonstrated that 6-MDS inhibited proliferation and induced Apoptosis and Autophagy. The Autophagy Inhibitor chloroquine and the silence of Atg5 augmented the effect of 6-MDS on promoting Apoptosis. Furthermore, 6-MDS suppressed the PI3K/Akt/mTOR signaling pathway, and the PI3K Inhibitor LY294002 enhanced these changes and promoted the 6-MDS pro-apoptotic and Autophagy effects. 6-MDS triggered the generation of Reactive Oxygen Species. The pretreatment with antioxidant N-acetyl-L-cysteine reversed the changes induced by 6-MDS, including increases in Apoptosis and Autophagy and inhibition of the PI3K/Akt/mTOR pathway. In conclusion, 6-MDS induces the Apoptosis and Autophagy of MCF-7 cells by ROS accumulation to suppress the PI3K/Akt/mTOR signaling pathway.