Gitogenin impedes tumorigenesis in hepatocellular carcinoma via NUAK2/NF-κB axis

Introduction Hepatocellular carcinoma (HCC), the most common primary malignant liver cancer, is associated with high incidence and mortality rates in China. Gitogenin (Gito), a saponin constituent in traditional Chinese medicine, exhibits significant pharmacological activity in promoting blood circulation and alleviating cerebral ischemia. However, the anti-HCC activity of Gito and its potential mechanism remain unknown. Methods A series of in vitro and in vivo experiments, including MTT assay, colony formation assay, immunofluorescence (IF) staining, Western blot, wound healing assay, Transwell migration and invasion assays, as well as patient-derived xenograft (PDX) mouse models of hepatocellular carcinoma (HCC), were performed to assess the anti-hepatocellular carcinoma effects of Gito and elucidate its underlying mechanisms. Results Our study found that Gito suppressed the growth and progression of HCC in vitro and in vivo by promoting autophagic processes and inhibiting epithelial-mesenchymal transition (EMT) and metastasis, likely through blocking the NUAK2/NF-κB axis. Autophagy inhibition reduced the inhibitory effects of Gito on the EMT, invasion, and migration of HCC cells. Additionally, NUAK2 overexpression weakened the inhibitory effect of Gito on the EMT, migration, and invasion of HCC cells, and the NF-κB axis. Gito treatment altered the melting curve of NUAK2 both in vitro and in vivo, suggesting that Gito may function as a potential inhibitor of NUAK2. Conclusion Our research provides a scientific basis for Gito regulating autophagy, EMT, and the metastatic abilities of HCC cells by inhibiting the NUAK2/NF-κB axis, indicating that Gito may be a promising candidate drug for HCC treatment.