Amentoflavone Enhances the Anti-tumor Activity of Regorafenib by Promoting Apoptosis and Inhibiting NF-κB–mediated Metastasis in Hepatocellular Carcinoma

Background/Aim: Regorafenib is approved for advanced hepatocellular carcinoma (HCC), but its limited efficacy and resistance remain challenges. Amentoflavone, a natural biflavonoid, has shown anticancer activity in preclinical studies.

Materials and Methods: This study assessed the combined effects of amentoflavone and regorafenib using in vitro assays and a Huh7 xenograft mouse model.

Results: Amentoflavone significantly enhanced regorafenib-induced cytotoxicity, activating both extrinsic (caspase-8) and intrinsic (caspase-9) apoptotic pathways. The combination also inhibited migration, invasion, and angiogenesis by suppressing VEGF-A, MMP-2, and phosphorylated NF-κB. In vivo, tumor growth was delayed over 10-fold in the combination group versus control, with reduced tumor weight and no evidence of systemic toxicity based on H&E staining, body weight, and liver/kidney function tests.

Conclusion: Amentoflavone potentiates regorafenib’s anti-tumor effects in HCC by promoting apoptosis and blocking NF-κB-mediated metastatic signaling, supporting its use as a safe and effective adjuvant strategy.