Introduction: Triple-negative breast cancer (TNBC) is a highly aggressive subtype of breast cancer with poor prognosis and limited targeted therapies. The development of novel effective drugs is of great significance for TNBC treatment.
Objective: This study aims to clarify the molecular mechanisms through which isorhamnetin(ISO) restrains the malignant progression of TNBC, usingcomputational pharmacology prediction and empirical verification.
Methods: We systematically screened public online databases and identified 198 putative targets of isorhamnetin and 1,490 genes functionally associated with TNBC. Overlapping targets were obtained through Venn diagram analysis. By integrating protein–protein interaction network profiling, functional enrichment analysis, and molecular docking experiments, we predicted the key binding targets of isorhamnetin. We then validated the effects of isorhamnetin on cell proliferation, migration, and invasion in TNBC cell lines. Western blot and cellular thermal shift assays were employed to examine target protein expression and alterations in downstream signaling pathways.
Results: The combination of protein–protein interaction network profiling, functional enrichment analysis, and molecular docking experiments indicated that ISO binds epidermal growth factor receptor (EGFR) with high affinity. Our findings revealed that ISO strongly reduced the growth, motility, and infiltrative capacities of TNBC cells. Western blot and cellular thermal shift assays confirmed that ISO downregulates EGFR protein expression and modulates downstream mammalian target of rapamycin (mTOR) and MAPK/ERK kinase (MEK)/extracellular signalregulated kinase (ERK) signaling.
Conclusion: ISO suppresses TNBC progression by downregulating EGFR signaling, providing a theoretical basis for its clinical application.