Apigenin in TMG inhibits colorectal tumorigenesis by targeting GSK3β/APC/β-catenin signaling pathway

Colorectal cancer (CRC) is a malignant tumor with a high global incidence. Tianma granule (TMG) is a traditional Chinese medicine compound. However, its pharmacodynamic material basis and mechanism of action against CRC remained unclear. This study systematically analyzed the anti-CRC mechanism of TMG by integrating metabolomics, network pharmacology, and experimental validation. HPLC-MS identified the main active components of TMG. Core targets and pathways were predicted by network pharmacology. The binding affinity between components and targets was evaluated by molecular docking. In vitro experiments utilized the HCT116 and HCT-8 cells model, where the effects of TMG and its key component Apigenin on cell activity. Apigenin was identified as a key flavonoid component in TMG by metabolomics. Network pharmacology analysis indicated that TMG intervened in CRC through multi-target actions, with the Wnt/β-catenin pathway being the core regulatory pathway. In vitro experiments demonstrated that both TMG and Apigenin inhibited HCT116 and HCT-8 cell proliferation, promoted apoptosis, and weakened invasion and migration capabilities. Apigenin promotes β-catenin degradation and suppresses its nuclear translocation by upregulating GSK3β and APC expression. The ablation or mutation of GSK3β or APC abolishes this effect, whereas β-catenin knockdown alone recapitulates the anti-cancer effects of TMG and Apigenin, demonstrating that their actions are strictly dependent on β-catenin. Apigenin, as the core active ingredient of TMG, was demonstrated to target GSK3β/APC to promote β-catenin degradation and inhibit the activation of the Wnt/β-catenin pathway, thereby exerting anti-CRC effects.