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Molecular mechanisms of euplotin C-induced apoptosis: involvement of mitochondrial dysfunction, oxidative stress and proteases

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Abstract

The metabolite euplotin C (EC), isolated from the marine ciliate Euplotes crassus, is a powerful cytotoxic and pro-apoptotic agent in tumour cell lines. For instance, EC induces the rapid depletion of ryanodine Ca2+ stores, the release of cytochrome c from the mitochondria, and the activation of caspase-3, leading to apoptosis. The purpose of this study was to gain further insight into the mechanisms of EC-induced apoptosis in rat pheochromocytoma PC12 cells. We found that EC increases Bax/Bcl-2 ratio and that Bax is responsible of the EC-induced dissipation of the mitochondrial membrane potential (Δψm). In addition, EC induces the generation of reactive oxygene species (ROS) without involvement of p53. The inhibition of ROS generation prevents, at least in part, the pro-apoptotic effects of EC as well as the effects of EC on Bax, Δψm and intracellular free Ca2+, indicating a cross-talk between different pathways. However, definition of the effector cascade turns out to be more complex than expected and caspase-independent mechanisms, acting in parallel with caspases, should also be considered. Among them, EC increases the expression/activity of calpains downstream of ROS generation, although calpains seem to exert protective effects.

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Acknowledgments

This research has been developed within the framework of the project INTERREG III A Italy-France (Sardegna-Corsica-Toscana) and financially supported by the Province of Livorno (Italy). We are greatful to Prof. G. Casini (University of Tuscia, Viterbo, Italy) for revising the manuscript. We also wish to thank Dr. B. Antonsson (Serono Pharmaceutical Research Institute, Geneva, Switzerland) for providing us with the Bax channel inhibitor Bci1.

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Authors and Affiliations

  1. Department of Environmental Sciences, University of Tuscia, largo dell’Università snc, blocco D, Viterbo, 01100, Italy

    Davide Cervia

  2. Department of Biology, University of Pisa, Pisa, 56127, Italy

    Davide Cervia, Mercedes Garcia-Gil, Elisa Simonetti, Graziano Di Giuseppe, Paola Bagnoli & Fernando Dini

  3. Department of Physics, Laboratory of Bioorganic Chemistry, University of Trento, Trento, 38050, Italy

    Graziano Guella

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  1. Davide Cervia
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  2. Mercedes Garcia-Gil
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  3. Elisa Simonetti
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  4. Graziano Di Giuseppe
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  5. Graziano Guella
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  7. Fernando Dini
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Correspondence to Davide Cervia.

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D. Cervia and M. Garcia-Gil equally contributed to the work.

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Cervia, D., Garcia-Gil, M., Simonetti, E. et al. Molecular mechanisms of euplotin C-induced apoptosis: involvement of mitochondrial dysfunction, oxidative stress and proteases. Apoptosis 12, 1349–1363 (2007). https://doi.org/10.1007/s10495-007-0075-7

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