Alharthy_2024_Drug.Chem.Toxicol__1

Anticancer medications cause anemia in patients through ill-defined mechanisms, including hemolysis and eryptosis. Although alpha-linolenic acid (ALA) possesses anticancer properties against a variety of cancer cells, there is a dearth of evidence regarding how it modulates red blood cell (RBC) physiology. RBCs from healthy donors were subjected to anticancer concentrations of ALA (2.5, 5, 10, 20, 40, 80, and 100 microM) at 37 degreesC for 24 h, and colorimetric tests were used to determine hemolysis and acetylcholinesterase (AChE) activity. Meanwhile, flow cytometry was employed to identify eryptotic cells using annexin-V-FITC and forward scatter (FSC), Fluo4/AM to detect Ca(2+), and H(2)DCFDA to assess oxidative stress. ALA significantly increased hemolysis and eryptosis in a concentration-dependent manner, along with elevated Fluo4 and DCF fluorescence, and erythrocyte sedimentation rate, and reduced FSC and AChE activity. Moreover, the addition of D4476, necrosulfonamide, melatonin, isosmotic urea, and polyethylene glycol 8000 - but not sucrose - significantly inhibited ALA toxicity. In conclusion, ALA stimulates hemolysis and eryptosis through Ca(2+) buildup, oxidative stress, anticholinesterase activity, casein kinase 1alpha (CK1alpha), and mixed lineage kinase domain-like protein (MLKL). The anticancer activity of ALA may be potentiated by the use of Ca(2+) channel blockers and chelators, antioxidants, and CK1alpha and MLKL inhibitors to ameliorate its toxicity to RBCs.