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.

中文翻译：

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6-甲氧基二氢血根碱通过积累 ROS 抑制 PI3K/AKT/mTOR 信号通路诱导乳腺癌 MCF-7 细胞凋亡和自噬

6-Methoxydihydrosanguinarine (6-MDS) 是一种从Hylomecon japonica中提取的天然苯并菲啶生物碱（Thunb。）Prantl。首次探讨6-MDS对乳腺癌的作用及机制。采用网络药理学、分子对接和分子动力学模拟技术，确定了6-MDS在乳腺癌中的潜在靶点和通路。此外，还进行了细胞增殖、凋亡和蛋白质印迹试验，以研究 6-MDS 对 MCF-7 细胞的影响。网络药理学、分子对接和分子动力学模拟结果证实了6-MDS通过PI3K/AKT/mTOR信号通路发挥抗乳腺癌作用。此外，功能实验结果表明6-MDS抑制增殖并诱导细胞凋亡和自噬。自噬抑制剂氯喹和 Atg5 的沉默增强了 6-MDS 促进细胞凋亡的作用。此外，6-MDS 抑制 PI3K/AKT/mTOR 信号通路，PI3K 抑制剂 LY294002 增强了这些变化，促进了 6-MDS 的促凋亡和自噬作用。6-MDS 引发活性氧的产生。用抗氧化剂 N-乙酰基-L-半胱氨酸预处理可逆转 6-MDS 诱导的变化，包括细胞凋亡和自噬的增加以及 PI3K/AKT/mTOR 通路的抑制。总之，6-MDS 通过 ROS 积累抑制 PI3K/AKT/mTOR 信号通路，诱导 MCF-7 细胞凋亡和自噬。用抗氧化剂 N-乙酰基-L-半胱氨酸预处理可逆转 6-MDS 诱导的变化，包括细胞凋亡和自噬的增加以及 PI3K/AKT/mTOR 通路的抑制。总之，6-MDS 通过 ROS 积累抑制 PI3K/AKT/mTOR 信号通路，诱导 MCF-7 细胞凋亡和自噬。用抗氧化剂 N-乙酰基-L-半胱氨酸预处理可逆转 6-MDS 诱导的变化，包括细胞凋亡和自噬的增加以及 PI3K/AKT/mTOR 通路的抑制。总之，6-MDS 通过 ROS 积累抑制 PI3K/AKT/mTOR 信号通路，诱导 MCF-7 细胞凋亡和自噬。