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Digestive system cancers,including liver,gastric,colon,esophageal and pancreatic cancers,are the leading cause of cancers with high morbidity and mortality,and the question of their clinical treatment is still open.Previous studies have indicated that Ziyuglycoside Ⅱ (ZYG Ⅱ),the major bioactive ingredient extract from Sanguisorba officinalis L.,significantly inhibits the growth of various cancer cells.However,the selective anti-tumor effects of ZYG Ⅱ against digestive system cancers are not systemically investigated.In this study,we reported the anti-cancer effect of ZYG Ⅱ on esophageal cancer cells (OE21),cholangiocarcinoma cells (HuCCT1),gastric cancer cells (BGC-823),liver cancer cells (HepG2),human colonic cancer cells (HCT116),and pancreatic cancer cells (PANC-1).We also found that ZYG Ⅱ induced cell cycle arrest,oxidative stress and mitochondrial apoptosis.Network pharmacology analysis suggested that UBC,EGFR and IKBKG are predicted targets of ZYG Ⅱ.EGFR signaling was suggested as the critical pathway underlying the anti-cancer effects ofZYG Ⅱ and both docking simulation and western blot analysis demonstrated that ZYG Ⅱ was a potential EGFR inhibitor.Furthermore,our results showed synergistic inhibitory effects of ZYG Ⅱ and chemotherapy 5-FU on the growth of cancer cells.In summary,ZYG Ⅱ are effective anti-tumor agents against digestive cancers.Further systemic evaluation of the anti-cancer activities in vitro and in vivo and characterization of underlying mechanism will promote the development of novel supplementary therapeutic strategies based on ZYG Ⅱ for the treatment of digestive system cancers.