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Non-apoptotic programmed cell death induced by a copper(II) complex in human fibrosarcoma cells

Histochemistry and Cell Biology volume 126, pages 473–482 (2006)Cite this article

Abstract

A0, a Cu(II) thioxotriazole complex, produces severe cytotoxic effects on HT1080 human fibrosarcoma cells with a potency comparable to that exhibited by cisplatin. A0 induced a characteristic series of changes, hallmarked by the formation of eosin- and Sudan Black-B-negative vacuoles. No evidence of nuclear fragmentation or caspase-3 activation was detected in cells treated with A0 which, rather, inhibited cisplatin-stimulated caspase-3 activity. Membrane functional integrity, assessed with calcein and propidium iodide, was spared until the late stages of the death process induced by the copper complex. Vacuoles were negative to the autophagy marker monodansylcadaverine and their formation was not blocked by 3-methyladenine, an inhibitor of autophagic processes. Negativity to the extracellular marker pyranine excluded vacuole derivation from the extracellular fluid. Ultrastructural analysis indicated that A0 caused the appearance of many electronlight cytoplasmic vesicles, possibly related to the endoplasmic reticulum, which progressively enlarge and coalesce to form large vacuolar structures that eventually fill the cytoplasm. It is concluded that A0 triggers a non-apoptotic, type 3B programmed cell death (Clarke in Anat Embryol (Berl) 181:195–213, 1990), characterized by an extensive cytoplasmic vacuolization. This peculiar cytotoxicity pattern may render the employment of A0 to be of particular interest in apoptosis-resistant cell models.

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Acknowledgments

S. T. is the recipient of a grant from Lega italiana per la lotta contro i tumori. This research has been partially funded by PRIN 2004059078. The confocal apparatus is a facility of CIM, University of Parma. Help from Dr. Cecilia Bozzetti in cytofluorimetric analysis, Dr. Paolo Govoni for histochemistry, and Dr. Silvana Belletti for technical assistance is gratefully acknowledged. We are also grateful to Prof. Renato Scandroglio and Prof. Gian C. Gazzola for discussions and comments.

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Affiliations

  1. Unit of General and Clinical Pathology, Department of Experimental Medicine, University of Parma, via Volturno, 39-43100, Parma, Italy

    S. Tardito, O. Bussolati, F. Gaccioli & R. Franchi-Gazzola

  2. Unit of Histology, Department of Experimental Medicine, University of Parma, via Volturno, 39-43100, Parma, Italy

    R. Gatti, S. Guizzardi & J. Uggeri

  3. Department of General and Inorganic, Analytical, and Physical Chemistry, University of Parma, Viale delle Scienze, 43100, Parma, Italy

    L. Marchiò & M. Lanfranchi

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  1. S. Tardito
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  2. O. Bussolati
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  3. F. Gaccioli
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  4. R. Gatti
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  5. S. Guizzardi
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  6. J. Uggeri
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  7. L. Marchiò
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  8. M. Lanfranchi
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  9. R. Franchi-Gazzola
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Correspondence to R. Franchi-Gazzola.

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Tardito, S., Bussolati, O., Gaccioli, F. et al. Non-apoptotic programmed cell death induced by a copper(II) complex in human fibrosarcoma cells. Histochem Cell Biol 126, 473–482 (2006). https://doi.org/10.1007/s00418-006-0183-4

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  • DOI: https://doi.org/10.1007/s00418-006-0183-4

Keywords

  • Programmed cell death
  • Vacuoles
  • Fibrosarcoma
  • Calcein
  • Cisplatin
  • Copper complex