Exploring the Mechanism of *Cimicifugae Rhizoma* against Breast Cancer via the EGFR‑MEK‑ERK Signaling Axis Based on Network Pharmacology

It integrates three types of data: network pharmacology prediction, molecular docking dynamics simulation, and 4T1 cell in vitro experiment. It is used to explore the molecular mechanism of Cimicifuga's intervention in breast cancer through the EGFR ‑ MEK ‑ ERK signal axis, which can provide an experimental and computational basis for mining the active ingredients of Cimicifuga. Based on TCMSP screening of active ingredients in sesame (OB ≥ 30%, DL ≥ 0.18), combined with SwissTarget Prediction to predict targets; GeneCards and OMIM acquire breast cancer targets; Venny 2.1 obtains drug disease intersection targets; STRING and Cytoscape 3.10.0 were used to construct a PPI network and screen for core targets; DAVID completes GO/KEGG enrichment.

AutoDock 4.2.6 carries out docking between active ingredients and core target molecules; Gromacs2025 completed a 100 ns molecular dynamics simulation of the coumarin-MMP9 complex, calculating parameters such as RMSD, RMSF, and MM-PBSA binding free energy. The extract of Cimicifuga was extracted with 78% ethanol, and 4T1 mouse breast cancer cells were used as the model; Using SRB method to detect cell proliferation and calculate IC ₅₀; Scratches and Transwell experiments were used to detect cell migration; Western blot was used to detect the expression of MMP9 and EGFR MEK ERK pathway proteins; Set up 3 biological replicates per group. Excel completes target sorting and deduplication; Cytoscape conducts network topology analysis; Gromacs tool analyzes simulated trajectories; ImageJ quantifies the grayscale of cell images and protein bands; GraphPad Prism was used for statistical analysis, and the results were expressed as \ (\ bar x \ pm s \)\ (P<0.05) indicates a statistically significant difference. database TCMSP、SwissTargetPrediction、UniProt、GeneCards、OMIM、STRING、DAVID、RCSB‑PDB、PubChem、Venny2.1。 Software: Cytoscape3.10.0、AutoDock4.2.6、PyMOL2.2.0、Gromacs2025、ImageJ、GraphPad Prism。 Instruments: Cell culture incubator, fluorescence inverted microscope, enzyme-linked immunosorbent assay reader, freeze centrifuge, electrophoresis and chemiluminescence imaging system, vacuum freeze dryer, etc.