The marine toxin palytoxin induces necrotic death in HaCaT cells through a rapid mitochondrial damage

- PLTX induces an early and irreversible necrotic cell death.
- Apoptosis occurrence was excluded.
- Necrosis is related to an early mitochondrial depolarization already after 5 min.
- Mitochondrial damage is strictly dependent on ionic imbalance.

Palytoxin (PLTX) is one of the most toxic algal biotoxin known so far. It transforms the Na+/K+-ATPase into a cationic channel inducing a massive intracellular Na+ influx. However, from a mechanistic point of view, the features and the intracellular pathways leading to PLTX-induced cell death are still not completely characterized. This study on skin HaCaT keratinocytes demonstrates that PLTX induces necrosis since propidium iodide uptake was observed already after 1 h toxin exposure, an effect that was not lowered by toxin removal. Furthermore, necrotic-like morphological alterations were evidenced by confocal microscopy. Apoptosis occurrence was excluded since no caspases 3/7, caspase 8, and caspase 9 activation as well as no apoptotic bodies formation were recorded. Necrosis was preceded by a very early mitochondrial damage as indicated by JC-1 fluorescence shift, recorded already after 5 min toxin exposure. This shift was totally abolished when Na+ and Ca2+ ions were withdrawn from culture medium, whereas cyclosporine-A was ineffective, excluding the occurrence of a controlled biochemical response.These results clearly establish necrosis as the primary mechanism for PLTX-induced cell death in HaCaT cells. The rapidity of mitochondrial damage and the consequent irreversible necrosis rise serious concerns about the very fast onset of PLTX toxic effects.