Based on network pharmacology and experiments to investigate the inhibitory effect of Licoricesaponin G2 on lung cancer tumor growth.

ETHNOPHARMACOLOGICAL RELEVANCE: Licoricesaponin G2 (LG2) is an active component in Licorice, which is a widely used traditional Chinese herbal medicine with antioxidant, anti-inflammatory, anti-cancer, and antiviral properties. LG2 can inhibit pneumonia, lung injury, and pulmonary fibrosis. However, the activity of LG2 against lung cancer and its mechanisms of action remain unclear. AIM OF THIS STUDY: This study aimed to elucidate the molecular mechanisms through which LG2 inhibits the progression of lung cancer, thereby establishing a theoretical framework for the development of novel targeted therapeutics for lung cancer. METHODS: The in vitro antitumor activity of LG2 was assessed using tumor sphere formation, colony formation, and 3D cell culture models, while its in vivo efficacy was confirmed in a subcutaneous xenograft model in nude mice. Potential target genes were identified by integrating data from Prediction, SwissTargetPrediction, GeneCards, and DisGeNET, followed by intersection with lung cancer-related targets. Network pharmacology analyses-including GO and KEGG enrichment, Reactome pathway analysis, and molecular docking-were employed to investigate underlying mechanisms. Key pathways and targets were subsequently validated through Western blotting, immunohistochemistry, and immunofluorescence. RESULTS: LG2 markedly inhibited lung cancer cell sphere and colony formation and significantly suppressed tumor growth in vivo. Integrated analysis combining network pharmacology, molecular docking, and experimental validation demonstrated that the principal targets of Licoricesaponin G2 are predominantly enriched in the PI3K/AKT signaling pathway. Ten critical targets were identified (STAT3, JUN, MMP9, MTOR, FGF2, PIK3R1, KDR, NFKB1, PDGFRB, TLR4). In addition, Licoricesaponin G2 substantially downregulated the protein expression of cancer stem cell markers CD44 and CD133, and inhibited the expression of ferroptosis marker proteins SLC7A11 and GPX4. CONCLUSION: LG2 mainly exerts anti-tumor effect on lung cancer by inhibiting the PI3K/AKT signaling pathway, regulating tumor stemness, and ferroptosis. These findings provide a mechanistic rationale and potential therapeutic strategy for the application of LG2 in lung cancer treatment.