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Two coffins and a funeral: early or late caspase activation determines two types of apoptosis induced by DNA damaging agents

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Abstract

Cell cytoskeleton makes profound changes during apoptosis including the organization of an Apoptotic Microtubule Network (AMN). AMN forms a cortical structure which plays an important role in preserving plasma membrane integrity during apoptosis. Here, we examined the cytoskeleton rearrangements during apoptosis induced by camptothecin (CPT), a topoisomerase I inhibitor, in human H460 and porcine LLCPK-1α cells. Using fixed and living cell imaging, we showed that CPT induced two dose- and cell cycle-dependent types of apoptosis characterized by different cytoskeleton reorganizations, time-dependent caspase activation and final apoptotic cell morphology. In the one referred as “slow” (~h) or round-shaped, apoptosis was characterized by a slow contraction of the actinomyosin ring and late caspase activation. In “slow” apoptosis the γ-tubulin complexes were not disorganized and microtubules were not depolymerized at early stages. In contrast, “fast” (~min) or irregular-shaped apoptosis was characterized by early caspase activation followed by full contraction of the actinomyosin ring. In fast apoptosis γ-tubulin complexes were disorganized and microtubules were initially depolymerized. However, after actinomyosin contraction, microtubules were reformed adopting a cortical but irregular disposition near plasma membrane. In addition to distinctive cytoskeleton reorganization kinetics, round and irregular-shaped apoptosis showed different biological properties with respect to AMN maintenance, plasma membrane integrity and phagocytes response. Our results suggest that the knowledge and modulation of the type of apoptosis promoted by genotoxic agents may be important for deciding a better therapeutic option and predicting the immune response in cancer treatment.

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Abbreviations

AMN:

Apoptotic microtubule network

CPT:

Camptothecin

CYTO:

Cytochalasin

FUCCI:

Fluorescent ubiquitination-based cell cycle indicator

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

MLC:

Myosin light chain

LDH:

Lactic dehydrogenase

LPA:

Lysophosphatidic acid

γTuRCs:

γ-tubulin ring complexes.

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Acknowledgements

This work was supported by FIS PI13/00129 grant, Instituto de Salud Carlos III, Spain and Fondo Europeo de Desarrollo Regional (FEDER-Unión Europea), Proyecto de Investigación de Excelencia de la Junta de Andalucía CTS-5725, and by AEPMI (Asociación de Enfermos de Patología Mitocondrial) and ENACH (Asociación de Enfermos de Neurodegeneración con Acumulación Cerebral de Hierro).

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    Authors and Affiliations

    1. Centro Andaluz de Biología del Desarrollo (CABD), and Centro de Investigación Biomédica en Red: Enfermedades Raras, Instituto de Salud Carlos III, Universidad Pablo de Olavide-Consejo Superior de Investigaciones Científicas, Carretera de Utrera Km 1, 41013, Sevilla, Spain

      Manuel Oropesa-Ávila, Patricia de la Cruz-Ojeda, Jesús Porcuna, Marina Villanueva-Paz, Alejandro Fernández-Vega, Mario de la Mata, Isabel de Lavera, Juan Miguel Suarez Rivero, Raquel Luzón–Hidalgo, Mónica Álvarez-Córdoba, David Cotán & José A. Sánchez-Alcázar

    2. Sistemas Físicos, Químicos y Naturales-Universidad Pablo de Olavide, 41013, Sevilla, Spain

      Ana Paula Zaderenko

    3. Facultad de Odontología, Universidad de Sevilla, 41009, Sevilla, Spain

      Mario D. Cordero

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    1. Manuel Oropesa-Ávila
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    Correspondence to José A. Sánchez-Alcázar.

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    Manuel Oropesa-Ávila and Patricia de la Cruz-Ojeda have contributed equally to this work.

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    Oropesa-Ávila, M., de la Cruz-Ojeda, P., Porcuna, J. et al. Two coffins and a funeral: early or late caspase activation determines two types of apoptosis induced by DNA damaging agents. Apoptosis 22, 421–436 (2017). https://doi.org/10.1007/s10495-016-1337-z

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