Andrographolide Induces Ferroptosis-associated Cytotoxicity and PUFA/MUFA Phospholipid Remodeling in Plasma Cell Neoplasm Cell Lines.

BACKGROUND/AIM: Andrographolide (Andro), a diterpene lactone from Andrographis paniculata, induces reactive oxygen species (ROS)-dependent apoptosis in hematologic malignancy cells. This study investigated whether Andro also induces ferroptosis-associated cytotoxicity in plasma cell neoplasm cell lines. MATERIALS AND METHODS: H929 and ARH77 cells were treated with Andro in the presence or absence of Ferrostatin-1 (Fer-1). Cell viability, ROS generation, Annexin V positivity, intracellular iron, lipid peroxidation, FACL4/GPX4 expression, nuclear factor kappa B (NF-κB)/cyclooxygenase-2 (COX-2) signaling, and lipidomic changes were examined. RESULTS: Andro induced dose-dependent cytotoxicity and increased ROS generation and Annexin V positivity, while Fer-1 partially restored Andro-induced loss of cell viability. Both cell lines showed elevated basal intracellular iron levels. Andro also induced Fer-1-sensitive lipid peroxidation, increased FACL4, decreased GPX4, and promoted lipid remodeling characterized primarily by reduced monounsaturated fatty acid (MUFA) abundance, resulting in an increased polyunsaturated fatty acid (PUFA)/MUFA ratio. In addition, Andro significantly decreased nuclear NF-κB p65 levels and COX-2 expression in both cell lines. CONCLUSION: Andro induces a ferroptosis-prone lipid state characterized by Fer-1-sensitive lipid peroxidation, FACL4 up-regulation, GPX4 down-regulation, and alterations in the PUFA/MUFA phospholipid balance. Suppression of the nuclear NF-κB/COX-2 axis may be associated with this lipid remodeling, although its causal role requires further investigation.