〔Abstract〕 Objective To investigate the effects of indirubatin derivative E804 on proliferation and migration of non-small cell lung cancer(NSCLC) A549 cells, and to elucidate the possible mechanism of Nrf2-HO-1/GPX4 pathway. Methods Lung cancer A549 cells were used as the cell model. The proliferation and migration of different specific inhibitors(Nec-1, CQ, Z-VAD, DFO, Fer-1 and Lip-1) in 0, 10 μmol/L E804 and 10 μmol/L E804+ groups were observed by MTT and cell scratch assay. The contents of reactive oxygen species(ROS) were detected by DCFH-DA fluorescence probe method, the contents of Fe2+were detected by colorimetric method, the contents of reduced glutathione(GSH) were detected by spectrophotometry, and the contents of malondialdehyde(MDA) were detected by micromethod. The expression levels of SLC7A11, Transferrin, GPX4, SLC40A1, Nrf2 and HO-1 were detected by Western blot in cells of 0, 2.5, 5 and 10 μmol/L E804 groups. Results Compared with the control group(0 μmol/L E804), 2.5, 5 and 10 μmol/L E804 significantly increased intracellular ROS, Fe2+and MDA levels, and decreased intracellular GSH content(P<0.01). Meanwhile, the expression levels of SLC7A11, GPX4, SLC40A1, Nrf2 and HO-1 significantly decreased(P<0.01), and the expression level of Transferrin increased(P<0.05). Compared with the 10 μmol/L E804 group alone, the apoptosis inhibitor(Z-VAD) group and the ferroptosis inhibitor(DFO, Fer-1 and Lip-1) group could significantly reverse the inhibition of proliferation and migration of A549 cells by 10 μmol/L E804(P<0.01). Conclution E804 can induce ferroptosis and inhibit the proliferation and migration of A549 cells, which may be related to the inhibition of Nrf2-HO-1/GPX4 pathway.