AMPD2 as a novel regulator of ferroptosis

Abstract Purpose Ferroptosis is a newly discovered programmed cell death, but its molecular mechanism remains largely unknown. AMPD2 is a protein, which related to oxygen metabolism in non-small cell lung cancer (NSCLC). In this study, we aimed to clarify whether AMPD2 is a regulatory factor of ferroptosis, which may provide a new theoretical basis for inducing ferroptosis to treat tumors. Methods we constructed AMPD2 overexpression and knockdown cells lines using lentivirus packaging technique. Immunohistochemistry was used to detect the expression of AMPD2 in cancerous tissue and adjacent normal tissue. DCFH-DA probe was used to detect the changes of reactive oxygen species(ROS). MDA assay was used to detect the changes of lipid peroxidation. CCK-8 assay was used to detect cell viability. Correlations of AMPD2 with Ferroptosis-related signaling pathways were analyzed using GEPIA. Western blot assay was used to detect changes in Ferroptosis-related molecular markers and signaling pathway. Results AMPD2 is higher expressed in NSCLC patient’s tissues. The Overexpression of AMPD2 significantly reduces intracellular ROS and MDA in NSCLC cells, inhibits ferroptosis induced by RSL3 (ferroptosis activator), enhances the expression of p38, p-p38 and GPX4, whereas knockdown of AMPD2 reverse the results. Moreover, treatment with AMP will increase ferroptosis sensitivity medicated by AMPD2. Conclusions AMPD2 regulates cellular ferroptosis by modulating the level of AMP affecting the p38 pathway and GPX4.