Activation of AMPK by Sophoricoside Suppresses Primary Liver Cancer Progression In Vitro and In Vivo

Primary liver cancer (PLC), mainly hepatocellular carcinoma (HCC), remains a leading global cause of cancer mortality. Metabolic dysregulation is a hallmark of HCC progression. Sophoricoside (SOP), a flavonoid glycoside from Sophora japonica, exhibits anti-inflammatory and metabolic regulatory activities, yet its role in liver cancer is unclear. Here, we investigated the antitumor effects and underlying mechanisms of SOP in HCC. Huh7 and HepG2 cells were treated with increasing SOP concentrations to assess proliferation, migration, and invasion using CCK-8, colony formation, wound-healing, and Transwell assays. Transcriptomic profiling (RNA-seq) and pathway enrichment analyses were conducted, followed by RT-qPCR and Western blot validation. In vivo efficacy was evaluated in Huh7 xenografts (160 mg/kg, daily for 27 days). SOP inhibited proliferation in a dose-dependent manner (IC₅₀ ≈ 320 μM), reducing clonogenicity by ~65% (p < 0.01) and suppressing migration and invasion by 60%-70%. These effects correlated with decreased PCNA, Cyclin D1, and N-cadherin, and upregulated E-cadherin. RNA-seq revealed 1263 differentially expressed genes, with AMPK pathway activation and repression of G2M, E2F, and MYC signaling. SOP increased AMPK phosphorylation (Thr172; +2.5-fold at 300 μM) without affecting total AMPK. In vivo, SOP reduced tumor volume by ~60% and Ki67 expression while sustaining AMPK activation. Co-treatment with the AMPK inhibitor compound C abrogated these effects, confirming AMPK dependence. Collectively, SOP functions as a natural AMPK agonist that reprograms tumor metabolism and suppresses oncogenic signaling, highlighting its promise as a low-toxicity therapeutic candidate for liver cancer.