Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer with high mortality, representing a serious challenge to women’s health worldwide. Xiaoyao Sanjie Decoction (XYSJD) has shown promising anti-TNBC effects, yet the molecular mechanisms behind its therapeutic activity remain largely unexplored. This study aimed to investigate the efficacy of XYSJD and its active compound quercetin in TNBC and to elucidate the underlying mechanisms of action. The chemical composition of XYSJD was characterized using ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry (UHPLC-Q Exactive HFX-MS). Network pharmacology, bioinformatics, and molecular docking approaches were employed to predict potential molecular targets and pathways associated with XYSJD activity. In vitro, the effects of XYSJD on TNBC cell proliferation, migration, invasion, and apoptosis were assessed using CCK-8, EdU, wound healing, transwell, Hoechst-PI staining, and flow cytometry assays. Western blotting was performed to validate target pathway modulation. In vivo antitumor activity was evaluated using a TNBC xenograft mouse model.
Nine major constituents of XYSJD were identified, and 206 potential targets relevant to TNBC were predicted. Bioinformatics analyses highlighted the EZH2/AKT1 signaling axis as a key pathway involved in XYSJD’s anticancer activity, with apoptosis emerging as a primary biological process affected. Both XYSJD and quercetin significantly inhibited TNBC cell growth and induced apoptosis in vitro and in vivo. Mechanistically, quercetin suppressed TNBC progression by downregulating the EZH2/AKT1 pathway, whereas overexpression of EZH2 or AKT1, or activation of AKT via SC79, attenuated quercetin-induced apoptosis. XYSJD exhibits potent antitumor effects against TNBC, with quercetin acting as a principal bioactive compound that triggers apoptosis through inhibition of the EZH2/AKT1 pathway. These findings provide a mechanistic rationale for the potential clinical application of XYSJD in TNBC therapy.
