Xiao Chai Hu Tang-derived decoction (Tung-Yi Fang) suppresses triple negative breast cancer cells in vitro and in vivo via regulating EGFR/AXL-mediated signaling

Background Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. Xiao Chai Hu Tang (XCHT), a classical herbal formula, is prescribed as an adjuvant therapy in breast cancer care. However, the high sugar content of XCHT may influence its anticancer efficacy under modern experimental conditions. This study investigated the anti-TNBC effects of Tung-Yi Fang (TYF), a sugar-reduced XCHT-derived decoction, and elucidated its underlying mechanisms in vitro and in vivo . Methods TYF was chemically characterized by reverse-phase high-performance liquid chromatography. Its bioactivities were evaluated in TNBC cells using viability and colony formation assays, as well as migration/invasion, cell cycle, and apoptosis analyses. Mechanistic insights were investigated using receptor tyrosine kinase (RTK) arrays, Western blotting, and immunofluorescence analyses. The in vivo relevance of TYF was further assessed in an orthotopic 4T1-luciferase breast cancer mouse model. Results TYF selectively suppressed TNBC cell growth while exerting minimal effects on non-tumorigenic breast epithelial cells and fibroblasts. TYF induced G2/M cell cycle arrest and apoptotic responses. TYF impaired TNBC cell mobility through disruption of actin cytoskeletal organization and suppression of FAK/Src signaling. RTK profiling and downstream analyses revealed that TYF concurrently regulated EGFR- and AXL-associated signaling pathways, leading to attenuation of AKT, ERK, and STAT3 activation. TYF administration significantly reduced tumor growth and metastatic burden without detectable systemic toxicity in vivo . Conclusion TYF exhibits anti-TNBC activity through EGFR and AXL inhibition and suppression of metastatic potential. Its efficacy highlights the therapeutic potential of glucose-controlled herbal formulations in cancer management.