Advertisement

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

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.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 99

This is the net price. Taxes to be calculated in checkout.

References

  1. 1.

    Nabha SM, Mohammad RM, Dandashi MH, et al. (2002) Combretastatin-A4 prodrug induces mitotic catastrophe in chronic lymphocytic leukemia cell line independent of caspase activation and poly(ADP-ribose) polymerase cleavage. Clin Cancer Res 8:2735–741

  2. 2.

    Kanthou C, Greco O, Stratford A, Cook I, Knight R, Benzakour O, Tozer G (2004) The tubulin-binding agent combretastatin A-4-phosphate arrests endothelial cells in mitosis and induces mitotic cell death. Am J Pathol 165:1401–411

  3. 3.

    Castedo M, Perfettini JL, Roumier T, Andreau K, Medema R, Kroemer G (2004) Cell death by mitotic catastrophe: a molecular definition. Oncogene 23:2825–837

  4. 4.

    Nitta M, Kobayashi O, Honda S, et al. (2004) Spindle checkpoint function is required for mitotic catastrophe induced by DNA-damaging agents. Oncogene 23:6548–558

  5. 5.

    Musacchio A, Hardwick KG (2002) The spindle checkpoint: structural insights into dynamic signalling. Nat Rev Mol Cell Biol 3:731–41

  6. 6.

    Millband DN, Campbell L, Hardwick KG (2002) The awesome power of multiple model systems: interpreting the complex nature of spindle checkpoint signaling. Trends Cell Biol 12:205–09

  7. 7.

    Zamzami N, Kroemer G (2003) Apoptosis: mitochondrial membrane permeabilization—the (w)hole story? Curr Biol 13:R71–3

  8. 8.

    Henry-Mowatt J, Dive C, Martinou JC, James D (2004) Role of mitochondrial membrane permeabilization in apoptosis and cancer. Oncogene 23:2850–860

  9. 9.

    Liu X, Zou H, Slaughter C, Wang X (1997) DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 89:175–84

  10. 10.

    Sakahira H, Enari M, Nagata S (1998) Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391:96–9

  11. 11.

    Castedo M, Perfettini JL, Roumier T, et al. (2004) Mitotic catastrophe constitutes a special case of apoptosis whose suppression entails aneuploidy. Oncogene 23:4362–370

  12. 12.

    Swanson PE, Carroll SB, Zhang XF, Mackey MA (1995) Spontaneous premature chromosome condensation, micronucleus formation, and non-apoptotic cell death in heated HeLa S3 cells. Ultrastructural observations. Am J Pathol 146:963–71

  13. 13.

    Ianzini F, Mackey MA (1997) Spontaneous premature chromosome condensation and mitotic catastrophe following irradiation of HeLa S3 cells. Int J Radiat Biol 72:409–21

  14. 14.

    Roninson IB, Broude EV, Chang BD (2001) If not apoptosis, then what? Treatment-induced senescence and mitotic catastrophe in tumor cells. Drug Resist Updat 4:303–13

  15. 15.

    Broker LE, Huisman C, Ferreira CG, Rodriguez JA, Kruyt FA, Giaccone G (2002) Late activation of apoptotic pathways plays a negligible role in mediating the cytotoxic effects of discodermolide and epothilone B in non-small cell lung cancer cells. Cancer Res 62:4081–088

  16. 16.

    Huisman C, Ferreira CG, Broker LE, Rodriguez JA, Smit EF, Postmus PE, Kruyt FA, Giaccone G (2002) Paclitaxel triggers cell death primarily via caspase-independent routes in the non-small cell lung cancer cell line NCI-H460. Clin Cancer Res 8:596–06

  17. 17.

    Nabha SM, Mohammad RM, Dandashi MH, et al. (2002) Combretastatin-A4 prodrug induces mitotic catastrophe in chronic lymphocytic leukemia cell line independent of caspase activation and poly(ADP-ribose) polymerase cleavage. Clin Cancer Res 8:2735–741

  18. 18.

    Broker LE, Huisman C, Span SW, Rodriguez JA, Kruyt FA, Giaccone G (2004) Cathepsin B mediates caspase-independent cell death induced by microtubule stabilizing agents in non-small cell lung cancer cells. Cancer Res 64:27–0

  19. 19.

    Vitale I, Antoccia A, Crateri P, Leone S, Arancia G, Tanzarella C (2005) Caspase-independent apoptosis is activated by diazepam-induced mitotic failure in HeLa cells, but not in human primary fibroblasts. Apoptosis 10:909–20

  20. 20.

    Leist M, Jaattela M (2001) Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2:589–98

  21. 21.

    Jaattela M (2004) Multiple cell death pathways as regulators of tumour initiation and progression. Oncogene 23:2746–756

  22. 22.

    Kagawa S, Gu J, Honda T, et al. (2001) Deficiency of caspase-3 in MCF7 cells blocks Bax-mediated nuclear fragmentation but not cell death. Clin Cancer Res 7:1474–80

  23. 23.

    Guicciardi ME, Leist M, Gores GJ (2004) Lysosomes in cell death. Oncogene 23:2881–890

  24. 24.

    Simoni D, Romagnoli R, Baruchello, et al. (2006) Novel combretastatin analogues endowed with antitumor activity. J Med Chem 49:3143–152

  25. 25.

    Cossarizza A, Baccarani-Contri M, Kalashnikova G, Franceschi C (1993) A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lypophilic cation 5,5′6,6′tetrachloro-1,1′3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1). Biochem Biophys Res Commun 197:40–5

  26. 26.

    Meschini S, Marra M, Condello M, Calcabrini A, Federici E, Dupuis ML, Cianfriglia M, Arancia, G (2005) Voacamine, an alkaloid extracted from Peschiera Fuchsiaefolia, inhibits P-glycoprotein action in multidrug-resistance tumor cells. Int J Oncol 27:1597–603

  27. 27.

    Dignam JD, Lebovitz RM, Roeder RG (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 11:1475–489

  28. 28.

    Gregoli PA, Bondurant MC (1999) Function of caspases in regulating apoptosis caused by erythropoietin deprivation in erythroid progenitors. J Cell Physiol 178:133–43

  29. 29.

    Mollinedo F, Gajate C (2003) Microtubules, microtubule-interfering agents and apoptosis. Apoptosis 8:413–50

  30. 30.

    Weaver BA, Cleveland DW (2005) Decoding the links between mitosis, cancer, and chemotherapy: The mitotic checkpoint, adaptation, and cell death. Cancer Cell 8:7–2

  31. 31.

    Tao W, South VJ, et al. (2005) Induction of apoptosis by an inhibitor of the mitotic kinesin KSP requires both activation of the spindle assembly checkpoint and mitotic slippage. Cancer Cell 8:49–9

  32. 32.

    Hall LL, Th’ng JP, Guo XW, Teplitz RL, Bradbury EM (1996) A brief staurosporine treatment of mitotic cells triggers premature exit from mitosis and polyploid cell formation. Cancer Res 56:3551–559

  33. 33.

    Masuda A, Maeno K, Nakagawa T, Saito H, Takahashi T (2003) Association between mitotic spindle checkpoint impairment and susceptibility to the induction of apoptosis by anti-microtubule agents in human lung cancers. Am J Pathol 163:1109–116

  34. 34.

    Taylor SS, McKeon F (1997) Kinetochore localization of murine Bub1 is required for normal mitotic timing and checkpoint response to spindle damage. Cell 89:727–35

  35. 35.

    Bhalla KN (2003) Microtubule-targeted anticancer agents and apoptosis. Oncogene 22:9075–086

  36. 36.

    Janmey PA (1998) The cytoskeleton and cell signaling: component localization and mechanical coupling. Physiol Rev 78:763–81

  37. 37.

    Leist M, Jaattela M (2001) Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2:589–98

  38. 38.

    Broker LE, Kruyt FA, Giaccone G (2005) Cell death independent of caspases: a review. Clin Cancer Res 11:3155–162

  39. 39.

    Kroemer G, Martin SJ (2005) Caspase-independent cell death. Nat Med 11:725–30

  40. 40.

    Minn AJ, Boise LH, Thompson CB (1996) Expression of Bcl-xL and loss of p53 can cooperate to overcome a cell cycle checkpoint induced by mitotic spindle damage. Genes Dev 10:2621–631

  41. 41.

    Lanni JS, Jacks T (1998) Characterization of the p53-dependent postmitotic checkpoint following spindle disruption. Mol Cell Biol 18:1055–064

  42. 42.

    von Haefen C, Wieder T, Essmann F, Schulze-Osthoff K, Dorken B, Daniel PT (2003) Paclitaxel-induced apoptosis in BJAB cells proceeds via a death receptor-independent, caspases-3/-8-driven mitochondrial amplification loop. Oncogene 22:2236–247

  43. 43.

    Jordan MA, Wendell K, Gardiner S, Derry WB, Copp H, Wilson L (1996) Mitotic block induced in HeLa cells by low concentrations of paclitaxel (Taxol) results in abnormal mitotic exit and apoptotic cell death. Cancer Res 56(4):816–25

  44. 44.

    Ofir R, Seidman R, Rabinski T, Krup M, Yavelsky V, Weinstein Y, Wolfson M (2002) Taxol-induced apoptosis in human SKOV3 ovarian and MCF7 breast carcinoma cells is caspase-3 and caspase-9 independent. Cell Death Differ 9:636–42

Download references

Author information

Correspondence to Caterina Tanzarella.

Additional information

Ilio Vitale and Antonio Antoccia contribuited equally to this work.

Electronic supplementary material

avi file: 59 MB

avi file: 54 MB

avi file: 34 MB

avi file: 59 MB

avi file: 54 MB

avi file: 34 MB

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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 (2007) doi:10.1007/s10495-006-0491-0

Download citation

Keywords

  • CA-4
  • ST2151
  • Spindle assembly checkpoint
  • Mitotic catastrophe
  • Caspase-3