This study aims to investigate the anti-liver cancer mechanisms of rhein.Potential therapeutic targets of rhein were retrieved from the TCMSP and SwissTargetPrediction databases,while liver cancer-related genes were retrieved from the TTD,GeneCards,OMIM,and PharmGKB databases.A "rhein-target-disease" interaction network was constructed to identify key targets.The protein-protein interaction network was constructed and key targets were identified.GO and KEGG pathway enrichment analyses were performed to identify relevant signaling pathways,and molecular docking was conducted between rhein and the key targets.In vitro,CCK-8 and scratch wound healing assays were used to assess rhein’s effects on Hepatocellular Carcinoma cell proliferation and migration.In vivo,the inhibitory effect of rhein on subcutaneous xenograft tumor growth was evaluated in nude mice.RT-qPCR and Western blotting were employed to quantify mRNA and protein expression levels of key targets.A total of 103 potential anti-primary liver cancer targets of rhein were identified,including matrix metalloproteinase 9 (MMP9),MMP2,mitogen-activated protein kinase 8 (MAPK8),and tissue inhibitor of metalloproteinases 1 (TIMP1).GO and KEGG analyses revealed significant enrichment in the MAPK signaling pathway.Molecular docking demonstrated strong binding affinities between rhein and core targets associated with MAPK signaling and extracellular matrix (ECM) degradation.Functionally,rhein suppressed HepG2 cell proliferation and migration in vitro and inhibited subcutaneous tumor growth in vivo.Mechanistically,rhein downregulated MAPK8 mRNA and protein expression while concurrently reducing ECM degradation-related targets,including MMP2,MMP9,TIMP1,TIMP2 and TIMP3.Collectively,these findings suggest that rhein exerts anti-liver cancer effects by inhibiting the MAPK signaling pathway and attenuating ECM degradation,providing a mechanistic foundation for its therapeutic potential in liver cancer.