Ferroptosis is identified as a regulated cell death mediated by iron accumulation and lipid peroxidation. The disturbances of mitochondrial morphology and function have been shown in this process. Mitochondrial Lon peptidase 1 (LONP1) is one of the main multi-function enzymes in regulating the mitochondrial function and cytological stability. To evaluate whether LONP1 take a role in ferroptosis, we applied erastin to initiate the ferroptosis in human pancreatic ductal adenocarcinoma (PDAC) cells. Here we show that erastin triggers cell death in both of oncogenic RAS mutant PANC1 cells and wild KRAS BxPC3 cells and the expression of LONP1 was up-regulated in this process. Gene inhibition of LONP1 only negatively regulates erastin-induced cell death and the alterations of molecular indicators in PANC1 cells. Furthermore, we show that inhibition of LONP1 activates the Nrf2/Keap1 signal pathway and up-regulates the expression of GPX4, a key peroxidase in regulating ferroptosis. Together, our results uncover a previously unappreciated mechanism coupling LONP1 to ferroptosis. (C) 2019 Elsevier Inc. All rights reserved.