Mitophagy dynamics during oxidative to reductive stress shift potentiates arsenite-induced malignant transformation...

Oxidative stress hypothesis of carcinogenicity from arsenic exposure is well-researched, reductive stress is increasingly emerging, and particularly, how cells undergo malignant transformation during the redox stress switch remains unclear. Herein, long-term (0-35 passages), low-dose (1.0 μM) sodium arsenite (NaAsO₂) exposure induces redox stress transition from oxidative to reductive stress in immortalized human keratinocytes (HaCaT) cells, driving malignant transformation via nuclear factor erythroid-2 related factor 2 (NRF2)-mediated PTEN-induced putative kinase 1 (PINK1)-PARK2 encoded E3 ubiquitin ligase (Parkin)-dependent mitophagy dynamics. Initially, NaAsO₂ elevates reactive oxygen species (ROS) to induce oxidative stress, but prolonged exposure reduces ROS while increasing reducing equivalents to form reductive stress at the cellular and mitochondrial levels. Sustained NRF2 activation post-ROS stimulation initiates mitophagy with a progressive enhancement from 42.8 % to 96 % through the PINK1-Parkin pathway, which is confirmed by inhibition with 10.0 μM cyclosporin A (CsA) or NRF2 siRNA, and activation with 10.0 μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP). This study highlights NRF2 modulates the redox homeostasis in a spatiotemporal dynamic manner and mitophagy dynamics works as a core mechanism underlying arsenite-induced cell malignant transformation, offering potential developing preventive and therapeutic strategies targeting mitophagy.