NOXA‐mediated degradation of MCL1 and BCL2L1 causes apoptosis of daunorubicin‐treated human acute myeloid leukemia cells

Daunorubicin (DNR) is used clinically to treat acute myeloid leukemia (AML), while the signaling pathways associated with its cytotoxicity are not fully elucidated. Thus, we investigated the DNR‐induced death pathway in the human AML cell lines U937 and HL‐60. DNR‐induced apoptosis in U937 cells accompanied by downregulation of MCL1 and BCL2L1, upregulation of Phorbol‐12‐myristate‐13‐acetate‐induced protein 1 (NOXA), and mitochondrial depolarization. DNR induced NOX4‐mediated reactive reactive oxygen species (ROS) production, which in turn inactivated Akt and simultaneously activated p38 mitogen‐activated protein kinase (MAPK). Activated p38 MAPK and inactivated Akt coordinately increased GSK3β‐mediated cAMP response element‐binding protein (CREB) phosphorylation, which promoted NOXA transcription. NOXA upregulation critically increased the proteasomal degradation of MCL1 and BCL2L1. The same pathway was also responsible for the DNR‐induced death of HL‐60 cells. Restoration of MCL1 or BCL2L1 expression alleviated DNR‐induced mitochondrial depolarization and cell death. Furthermore, ABT‐199 (a BCL2 inhibitor) synergistically enhanced the cytotoxicity of DNR in AML cell lines. Notably, DNR‐induced DNA damage was not related to NOXA‐mediated degradation of MCL1 and BCL2L1. Collectively, these results indicate that the upregulation of NOXA expression through the NOX4‐ROS‐p38 MAPK‐GSK3β‐CREB axis results in the degradation of MCL1 and BCL2L1 in DNR‐treated U937 and HL‐60 cells. This signaling pathway may provide insights into the mechanism underlying DNR‐triggered apoptosis in AML cells. Daunorubicin (DNR)‐induced NOX4‐mediated ROS generation activates p38 MAPK and inactivates Akt, which activates GSK3β by phosphorylation of Y216 and dephosphorylation of S9. GSK3β‐mediated CREB phosphorylation promotes NOXA transcription in U937 and HL‐60 cells. NOXA upregulation elicits MCL1 and BCL2L1 degradation, resulting in the activation of caspase‐3 and apoptosis of U937 and HL‐60 cells.