Mechanistic Exploration of Shugan Jianpi Formula for Treating Triple-Negative Breast Cancer Under Chronic Stress: A Network Pharmacology-Guided Experimental Study.

PURPOSE: This study aimed to investigate the pharmacological mechanisms of Shugan Jianpi Formula (SGJPF) in treating TNBC using network pharmacology and molecular biology approaches. METHODS: HPLC/MS identified the key compounds in SGJPF. In vitro assays were performed on norepinephrine (NE)-stimulated MDA-MB-231 and SUM159PT cells to mimic triple-negative breast cancer (TNBC) under chronic psychological stress (CPS) and evaluate SGJPF's effects on cell proliferation, apoptosis, cell cycle, migration, and invasion. A TNBC mouse model exposed to CPS was used to assess SGJPF's influence on tumor growth. SGJPF's mechanisms were explored via network pharmacology and molecular docking, with target validation through Western blotting, immunohistochemistry, and immunofluorescence. RESULTS: HPLC/MS analysis identified 806 compounds in SGJPF, including flavonoids, polyphenols, saponins, polysaccharides, alkaloids, terpenoids, coumarins, organic acids, and glycosides. Network pharmacology and molecular docking analyses identified SRC, ERK (MAPK1), and STAT3 as pivotal targets underlying the anti-tumor effects of SGJPF in TNBC. Both in vitro and in vivo experiments confirmed that SGJPF exerts its therapeutic effects through the modulation of the SRC/ERK/STAT3 signaling axis. In vitro, SGJPF effectively inhibited TNBC cell proliferation, migration, and invasion, while promoting apoptosis in NE-stimulated cells. In a CPS-induced TNBC mouse model, SGJPF significantly alleviated tumor progression, further corroborating its potential as a novel therapeutic strategy for TNBC. CONCLUSION: This study highlights the potential of SGJPF as a therapeutic strategy for TNBC through its modulation of the SRC/ERK/STAT3 signaling axis, offering a robust foundation for further investigation into its clinical application.