OBJECTIVE: To investigate the mechanism by which Biejiajian Pill (BJJP) regulates ferroptosis in hepatocellular carcinoma (HCC) cells through the p62/Keap1/NRF2 pathway and to provide an experimental basis for its application in the prevention and treatment of HCC. METHODS: Huh7 HCC cells were divided into a normal control group, a BJJP drug serum group, an erastin (a ferroptosis inducer) group, a BJJP drug serum + erastin group, and BJJP drug serum + ferrostatin-1 (Fer-1) (a ferroptosis inhibitor) group. BJJP drug serum was prepared with animals treated with BJJP and CCK-8 assay was performed to determine the optimal concentration and duration of BJJP intervention. The levels of intracellular iron (Fe), reduced glutathione (GSH), lipid peroxides (MDA), and reactive oxygen species (ROS) were measured. Western blot was performed to determine the expression levels of FTH1, GPX4, xCT, SLC40A1, Keapl, p62, and NRF2. JC-1 staining was performed to measure mitochondrial membrane potential, and cell immunofluorescence was performed to determine the expression of p62 and Keap1. RESULTS: According to the CCK-8 assay results, the cell inhibition rate was highest when BJJP was administered at a high dose of 2.2 g/kg ( CONCLUSION: BJJP regulates ferroptosis in Huh7 HCC cells by inhibiting the p62/Keap1/NRF2 pathway, demonstrating potentials as a therapeutic agent for HCC.