The aim of this study was to investigate the mechanism of action of oroxin A in inhibiting the malignant behavior of hepatocellular carcinoma cells.Methods The hepatoma cell lines Hep3B and MHCC97-L were purchased.We set gradient concentration of oroxin A(0, 30, 60, 90, 120 μM), and the semi-inhibitory concentration (IC50) of oroxin A on hepatoma cells was determined by CCK-8 assay of cell viability.IC50 concentration was subsequently taken for subsequent experiments.The cells were grouped as Control and OA.Kits were taken to detect iron death-related indexes Fe2+ content, lipid reactive oxygen species (Lipid-ROS) content, glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD) activity, and cystine uptake levels using the Tri-Carb Liquid Scintillation Analyzer.We transfected pcDNA3.1-TXNDC5 into HCC cells induced by oroxin A (OA+oe-TXNDC5), with pcDNA3.1-NC as control (OA+oe-NC).The effect of overexpression of TXNDC5 on ferroptosis in HCC cells induced by oroxin A was examined.Resluts A concentration-dependent inhibition of hepatocellular carcinoma cell MHCC97-L and Hep3B viability was observed. The semi-inhibitory concentration of Hep3B was 30 μM.Treatment of Hep3B cells with oroxin A resulted in a significant increase in Fe2+ content, inhibition of cystine uptake, significant increase in Lipid-ROS content and MDA level, decrease in GSH content and SOD activity, which mean a induction of ferroptosis in Hep3B cells.Overexpression of TXNDC5 partially reversed the induction of ferroptosis by Oroxin A.Conclusion 〗Oroxin A induced ferroptosis in HCC cells by downregulating TXNDC5, thereby inhibiting the malignant behavior of hepatocellular carcinoma cells