Neratinib inhibits proliferation and promotes apoptosis of acute myeloid leukemia cells by activating autophagy‐dependent ferroptosis

Abstract Acute myeloid leukemia (AML) is a hematologic malignancy with increased lethality. We focused on elucidating the role of Neratinib, a tyrosine kinase inhibitor, in the progression of AML and identify the potential mechanisms. Upon the treatment of Neratinib, autophagy suppressor 3‐methyladenine (3‐MA) and ferroptosis stimulator Erastin, the viability and proliferation of HL‐60 cells were evaluated by cell counting kit‐8 and 5‐Ethynyl‐20‐Deoxyuridine staining assays. A flow cytometer was to observe cell cycle and apoptosis. Production of reactive oxygen species (ROS) was tested via 2,7‐dichlorodihydrofluorescein diacetate assay. Additionally, malondialdehyde (MDA) content and Fe 2+ activity were examined with commercial kits. LC3‐II expression was examined by using immunofluoresence staining. Western blot analysis ascertained the expression of proliferation, apoptosis, ferroptosis and autophagy‐associated proteins. It was noted that Neratinib notably mitigated cell viability and proliferation, cut down Ki67 and proliferating cell nuclear antigen expression. Moreover, Neratinib hindered cell cycle at G0/G1 phase whereas exacerbated apoptosis. ROS, MDA and Fe 2+ activities were elevated by Neratinib, coupled with the reduced glutathione peroxidase 4, ferritin heavy chain 1 expression and enhanced acyl‐CoA synthetase long‐chain family member 4 expression. Furthermore, Neratinib promoted autophagy of HL‐60 cells, evidenced by raised LC3‐II, ATG5, Beclin1 expression and lessened p62 expression. Importantly, 3‐MA eased the impacts of Neratinib on cell ferroptosis, proliferation and apoptosis, which were offset by further administration of Erastin. To conclude, Neratinib could suppress proliferation and promote apoptosis of HL‐60 cells through autophagy‐dependent ferroptosis.