Ginsenoside RG3 and cantharidin synergistically suppress the progression of hepatocellular carcinoma via targeting the PRMT1-SREBF1 axis-mediated lipid metabolism

Ginsenosides, such as ginsenoside RG3, demonstrate antitumor potential in hepatocellular carcinoma (HCC) and are often combined with cantharidin (CTD) in traditional Chinese medicine to achieve synergistic effects while mitigating CTD’s toxicity. However, the precise molecular mechanisms underlying this synergy remain elusive. The progression of HCC was assessed using a series of in vitro assays, including CCK-8 for cell viability, EdU staining for proliferation, wound healing for migration, and transwell assay for invasion. The antitumor efficacy and hepatotoxicity were assessed in animal models using mice, employing tumor volume measurement, histopathological analysis, and quantification of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) through ELISA. To decipher the underlying synergistic mechanisms, we employed an integrated approach of network pharmacology, RNA sequencing, and molecular docking. The expression of key targets was verified by RT-qPCR and western blotting, while the direct interaction between PRMT1 and SREBF1 was confirmed by co-immunoprecipitation (Co-IP). The RG3/CTD combination exhibited a potent synergistic antitumor effect, suppressing tumor proliferation, migration, and invasion more effectively than either agent alone. Mechanistically, the therapy dually modulated aberrant lipid metabolism by concurrently inhibiting the PI3K/AKT/mTOR signaling axis and PRMT1-mediated epigenetic regulation. We identified a novel direct interaction between PRMT1 and SREBF1. The binding of the CTD/RG3 complex disrupted this interaction, inhibiting PRMT1-mediated arginine methylation of SREBF1 and consequently downregulating SREBF1 expression and activity. Furthermore, RG3 significantly mitigated CTD-induced hepatotoxicity by maintaining hepatic serum ALT and AST levels, an effect likely mediated by the modulation of AKT, ACOX1, and ABCB1 pathways to reduce oxidative stress and restore metabolic homeostasis. Our findings establish a novel RG3-CTD regimen that concurrently enhances therapeutic efficacy and reduces hepatotoxicity through coordinated targeting of oncogenic signaling and metabolic reprogramming. This study provides a robust mechanistic foundation for the clinical translation of RG3/CTD combination therapy for HCC.