Introduction Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer lacking effective therapeutic options. Berberine (BBR), a natural isoquinoline alkaloid, has shown anti-tumor potential, but its systemic mechanism of action in TNBC has not been fully elucidated. Methods This study adopted an integrated strategy combining network pharmacology, transcriptomic analysis, molecular docking, molecular dynamics simulation, and multi-level in vitro experimental validation. Targets of BBR were predicted through multiple databases, TNBC-related genes were screened by integrating GEO and TCGA data, key targets and tumor microenvironment characteristics were analyzed using WGCNA, PPI networks, and single-cell sequencing data, and the biological effects of BBR were validated through molecular docking, dynamics simulation, and cellular experiments. Results A total of 182 common targets of BBR and TNBC were identified, with 12 genes, including SRC, STAT3, and EGFR, being identified as hub targets. Immune infiltration analysis indicated that BBR targets were associated with the tumor immune microenvironment. Single-cell sequencing showed that these targets are primarily enriched in tumor cells and macrophages, and molecular docking and dynamics simulations revealed that BBR has high affinity with SRC (−8.8 kcal/mol). In vitro experiments confirmed that BBR can concentration-dependently inhibit TNBC cell proliferation, induce apoptosis, increase ROS levels, and significantly reduce the proportion of CD133 + stem cells and tumor sphere formation capacity. Western blot analysis showed that BBR downregulated the activity of the p-PI3K/p-AKT and p-SRC/p-STAT3 pathways. Conclusion BBR inhibits TNBC cell proliferation, induces apoptosis, weakens tumor stemness, and may regulate the tumor immune microenvironment through multi-target, multi-pathway synergistic effects. This study systematically reveals the integrative mechanism of BBR against TNBC, providing a theoretical basis for its development as a multi-target natural anti-tumor drug.
Zhang et al. (Thu,) studied this question.