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The tubulin-depolymerising agent combretastatin-4 induces ectopic aster assembly and mitotic catastrophe in lung cancer cells H460

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Abstract

The relationship between microtubular dynamics, dismantling of pericentriolar components and induction of apoptosis was analysed after exposure of H460 non-small lung cancer cells to anti-mitotic drugs. The microtubule destabilising agent, combretastatin-A4 (CA-4) led to microtubular array disorganization, arrest in mitosis and abnormal metaphases, accompanied by the presence of numerous centrosome-independent “star-like” structures containing tubulin and aggregates of pericentrosomal matrix components like γ-tubulin, pericentrin and ninein, whereas the structural integrity of centrioles was not affected by treatment. On the contrary, in condition of prolonged exposure or high concentrations of CA-4 such aggregates never formed. Treatment with 7.5 nM CA-4, which produced a high frequency “star-like” aggregates, was accompanied by mitotic catastrophe commitment characterized by translocation of the proapoptotic Bim protein to mitochondria activation of caspases-3/9 and DNA fragmentation as a result of either prolonged metaphase arrest or attempt of cells to divide. Drug concentrations which fail to block cells at mitosis were also unable to activate apotosis. A detailed time-course analysis of cell cycle arrest and apoptosis indicated that after CA-4 washout the number of metaphases with “star-like” structures decreased as a function of time and arrested cells proceeded in anaphase. After 4 h, the multiple α- and γ-tubulin aggregates coalesced into two well-defined spindles in a bipolar mitotic spindle organization. Overall, our findings suggest that the maintenance of microtubular integrity plays a relevant role in stabilising the pericentriolar matrix, whose dismantling can be associated with apoptosis after exposure to microtubule depolymerising agents.

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Acknowledgments

Chiara Cenciarelli is a PhD fellow at the Department of Biology, Università Roma Tre. This work was partly financed by SIGMA-TAU.

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Authors and Affiliations

  1. Department of Biology, University “Roma Tre”, V.le Marconi 446, 00146, Rome, Italy

    Chiara Cenciarelli, Caterina Tanzarella, Ilio Vitale & Antonio Antoccia

  2. SIGMA-TAU Industrie Farmaceutiche Riunite S.p.A, Via Pontina km 30,400, 00040, Pomezia, Italy

    Claudio Pisano

  3. Department of Technology and Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy

    Pasqualina Crateri, Stefania Meschini & Giuseppe Arancia

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  1. Chiara Cenciarelli
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Correspondence to Antonio Antoccia.

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Cenciarelli, C., Tanzarella, C., Vitale, I. et al. The tubulin-depolymerising agent combretastatin-4 induces ectopic aster assembly and mitotic catastrophe in lung cancer cells H460. Apoptosis 13, 659–669 (2008). https://doi.org/10.1007/s10495-008-0200-2

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