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

Fen Liu,1,* Luning Li,2,* Yiming Zhang,3,* Jiaqi Zhang,2 Xinchen Tian,2 Dengtian Zhang,4 Ni Zhang,5 Tinghao Yan,5 Shulong Shi,6 Jianlin Wu,1,7 Shulong Jiang2,3,5 1College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250000, People’s Republic of China; 2Clinical Medical Laboratory Center, Jining First People’s Hospital, Shandong First Medical University, Jining, 272000, People’s Republic of China; 3College of First Clinical Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250000, People’s Republic of China; 4Department of Radiation, The 960th Hospital of the PLA Joint Logistics Support Force, Jinan, 250000, People’s Republic of China; 5Cheeloo College of Medicine, Shandong University, Jinan, 250000, People’s Republic of China; 6Department of Endocrinology, Jining First People’s Hospital, Shandong First Medical University, Jining, 272000, People’s Republic of China; 7Key Laboratory of Traditional Chinese Medicine Classical Theory, Ministry of Education, Shandong University of Traditional Chinese Medicine, Jinan, 250000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Shulong Jiang, Clinical Medical Laboratory Center, Jining First People’s Hospital, Shandong First Medical University, Jining, 272000, People’s Republic of China, Email [email protected] Shulong Shi, Department of Endocrinology, Jining First People’s Hospital, Shandong First Medical University, Jining, 272000, People’s Republic of China, Email [email protected]: 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.Keywords: Shugan Jianpi formula, triple-negative breast cancer, chronic stress, network pharmacology, SRC/ERK/STAT3 pathway