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Combretastatin CA-4 and combretastatin derivative induce mitotic catastrophe dependent on spindle checkpoint and caspase-3 activation in non-small cell lung cancer cells

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Abstract

Combretastatin A-4 (CA-4), a natural stilbenoid isolated from Combretum caffrum, is a new vascular targeting agent (VTA) known for its antitumor activity due to its anti-tubulin properties. We investigated the molecular mechanisms leading to cell death in non-small cell lung cancer H460 cells induced by natural (CA-4) and synthetic stilbenoids (ST2151) structurally related to CA-4. We found that both compounds induced depolymerization and rearrangement of spindle microtubules, as well as an increasingly aberrant organization of metaphase chromosomes in a dose- and time-dependent manner. Prolonged exposition to ST2151 led cells to organize multiple sites of tubulin repolymerization, whereas tubulin repolymerization was observed only after CA-4 washout. H460 cells were arrested at a pro-metaphase stage, with condensed chromosomes and a triggered spindle assembly checkpoint, as evaluated by kinetochore localization of Bub1 and Mad1 antibodies. Persistent checkpoint activation led to mitochondrial membrane permeabilization (MMP) alterations, cytochrome c release, activation of caspase-9 and -3, PARP cleavage and DNA fragmentation. On the other hand, caspase-2, and -8 were not activated by the drug treatment. The ability of cells to reassemble tubulin in the presence of an activated checkpoint may be responsible for ST2151-induced multinucleation, a recognized sign of mitotic catastrophe. In conclusion, we believe that discovery of new agents able to trigger mitotic catastrophe cell death as a result of mitotic block and prolonged spindle checkpoint activation is particularly worthwhile, considering that tumor cells have a high proliferative rate and mitotic failure occurs irrespective of p53 status.

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Authors and Affiliations

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

    Ilio Vitale, Antonio Antoccia, Chiara Cenciarelli & Caterina Tanzarella

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

    Pasqualina Crateri, Stefania Meschini & Giuseppe Arancia

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

    Claudio Pisano

Authors
  1. Ilio Vitale
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  2. Antonio Antoccia
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  3. Chiara Cenciarelli
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  4. Pasqualina Crateri
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  5. Stefania Meschini
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Correspondence to Caterina Tanzarella.

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Ilio Vitale and Antonio Antoccia contribuited equally to this work.

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Vitale, I., Antoccia, A., Cenciarelli, C. et al. Combretastatin CA-4 and combretastatin derivative induce mitotic catastrophe dependent on spindle checkpoint and caspase-3 activation in non-small cell lung cancer cells. Apoptosis 12, 155–166 (2007). https://doi.org/10.1007/s10495-006-0491-0

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  • DOI: https://doi.org/10.1007/s10495-006-0491-0

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