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This study systematically investigated the effects of Xihuang Pill on exosomes (Exo) secretion and how Exo pretreated with Xihuang Pill affect intercellular communication, thereby inhibiting the activity of triple-negative breast cancer cells. First, the inhibitory affected of Xihuang Pill on the activity and migration of 4T1 and MDA-MB-231 cells were verified using MTT, scratch assays, and crystal violet assays. Then, Exo and Exo secreted by cells pretreated with Xihuang Pill (XHP-Exo) were obtained by ultracentrifugation. The Exo were characterized by transmission electron microscopy, potential, particle size, and marker proteins. The inhibitory effects of XHP-Exo on the activity and migration of 4T1 and MDA-MB-231 cells were detected using crystal violet and scratch assays. The expression of programmed death ligand 1 (PD-L1), glutathione peroxidase 4 (GPX4), reactive oxygen species (ROS), and reduced glutathione (GSH) in cells was detected by Western blot and immunofluorescence. The chemical components of Xihuang Pill and XHP-Exo were analyzed by ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap mass spectrometry (UPLC-Q-Exactive-MS/MS). The results showed that Xihuang Pill could significantly inhibit the proliferation and migration of breast cancer cells, downregulate kidney and brain expressed adaptor protein (KIBRA) expression, and effectively reduce the secretion of exosomes from breast cancer cells. Furthermore, XHP-Exo induced ferroptosis in recipient tumor cells, characterized by increased intracellular ROS levels and decreased GSH content, accompanied by downregulation of PD-L1 expression. Component analysis revealed 19 common active chemical components in both Xihuang Pill and XHP-Exo. In summary, Xihuang Pill inhibits the secretion of exosomes from breast cancer cells, and component-containing exosomes derived from it induce ferroptosis in recipient cells and downregulate PD-L1 expression.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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