A combretastatin-mediated decrease in neutrophil concentration in peripheral blood and the impact on the anti-tumor activity of this drug in two different murine tumor models

PLoS One. 2014 Oct 9;9(10):e110091. doi: 10.1371/journal.pone.0110091. eCollection 2014.


The vascular disrupting agent combretastatin A-4 disodium phosphate (CA4P) induces fluctuations in peripheral blood neutrophil concentration. Because neutrophils have the potential to induce both vascular damage and angiogenesis we analyzed neutrophil involvement in the anti-tumoral effects of CA4P in C3H mammary carcinomas in CDF1 mice and in SCCVII squamous cell carcinomas in C3H/HeN mice. Flow cytometry analyses of peripheral blood before and up to 144 h after CA4P administration (25 and 250 mg/kg) revealed a decrease 1 h after treatment, followed by an early (3-6 h) and a late (>72 h) increase in the granulocyte concentration. We suggest that the early increase (3-6 h) in granulocyte concentration was caused by the initial decrease at 1 h and found that the late increase was associated with tumor size, and hence independent of CA4P. No alterations in neutrophil infiltration into the C3H tumor after CA4P treatment (25 and 250 mg/kg) were found. Correspondingly, neutrophil depletion in vivo, using an anti-neutrophil antibody, followed by CA4P treatment (25 mg/kg) did not increase the necrotic fraction in C3H tumors significantly. However, by increasing the CA4P dose to 250 mg/kg we found a significant increase of 359% in necrotic fraction when compared to neutrophil-depleted mice; in mice with no neutrophil depletion CA4P induced an 89% change indicating that the presence of neutrophils reduced the effect of CA4P. In contrast, neither CA4P nor 1A8 affected the necrotic fraction in the SCCVII tumors significantly. Hence, we suggest that the initial decrease in granulocyte concentration was caused by non-tumor-specific recruitment of neutrophils and that neutrophils may attenuate CA4P-mediated anti-tumor effect in some tumor models.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bibenzyls / administration & dosage
  • Carcinoma, Squamous Cell / blood*
  • Carcinoma, Squamous Cell / drug therapy
  • Carcinoma, Squamous Cell / pathology
  • Disease Models, Animal
  • Female
  • Flow Cytometry
  • Granulocytes / pathology
  • Humans
  • Mammary Neoplasms, Animal / blood*
  • Mammary Neoplasms, Animal / drug therapy
  • Mice
  • Necrosis / blood
  • Necrosis / drug therapy
  • Necrosis / pathology
  • Neovascularization, Pathologic / blood*
  • Neovascularization, Pathologic / drug therapy
  • Neovascularization, Pathologic / pathology
  • Neutrophils / drug effects*
  • Neutrophils / pathology


  • Bibenzyls
  • combretastatin

Grant support

Financial support for this study was provided by The Danish Cancer Society (www.cancer.dk), The Danish Council for Independent Research: Medical Sciences (www.fivu.dk), Radiumstationens research foundation, and The foundation in memory of Sigvald and Edith Anelise Celine Kristine Rasmussen, born Poulsen. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.