Tumor Biology

, Volume 36, Issue 11, pp 8499–8510 | Cite as

Combretastatin A4 phosphate treatment induces vasculogenic mimicry formation of W256 breast carcinoma tumor in vitro and in vivo

  • Nan Yao
  • Ke Ren
  • Cuihua Jiang
  • Meng Gao
  • Dejian Huang
  • Xiao Lu
  • Bin Lou
  • Fei Peng
  • Aizhen Yang
  • Xiaoning Wang
  • Yicheng Ni
  • Jian Zhang
Research Article


The purpose of this study was to investigate the effect of combretastatin A4 phosphate (CA4P) on vasculogenic mimicry (VM) channel formation in vitro and in vivo after a single-dose treatment and the underlying mechanism involved in supporting VM. In vitro model of three-dimensional cultures was used to test the effect of CA4P on the tube formation of Walker 256 cells. Western blot analysis was conducted to assess the expression of hypoxia-inducible factor (HIF)-1α and VM-associated markers. W256 tumor-bearing rat model was established to demonstrate the effect of CA4P on VM formation and tumor hypoxia by double staining and a hypoxic marker pimonidazole. Anti-tumor efficacy of CA4P treatment was evaluated by tumor growth curve. Under hypoxic conditions for 48 h in vitro, W256 cells formed VM network associated with increased expression of VM markers. Pretreatment with CA4P did not influence the amount of VM in 3-D culture as well as the expression of these key molecules. In vivo, W256 tumors showed marked intratumoral hypoxia after CA4P treatment, accompanied by increased VM formation. CA4P exhibited only a delay in tumor growth within 2 days but rapid tumor regrowth afterward. VM density was positively related to tumor volume and tumor weight at day 8. CA4P causes hypoxia which induces VM formation in W256 tumors through HIF-1α/EphA2/PI3K/matrix metalloproteinase (MMP) signaling pathway, resulting in the consequent regrowth of the damaged tumor.


Vasculogenic mimicry Combretastatin A4 phosphate (CA4P) Hypoxia Tumor regrowth 



This work was partially supported by the grants awarded by Jiangsu Province Natural Science Foundation (No. BK2012775), the Key Scientific and Technological Project of Changzhou Health Bureau (No. ZD201404), the National Natural Science Foundation of China (No. 81473120), hospital foundation of Jiangsu Branch of China Academy of Chinese Medical Science (JSBY1308), and Changzhou Siyao foundation of hospital pharmacy (2014YX011).

Conflicts of interest



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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Nan Yao
    • 1
  • Ke Ren
    • 2
    • 3
  • Cuihua Jiang
    • 1
  • Meng Gao
    • 1
  • Dejian Huang
    • 1
  • Xiao Lu
    • 4
  • Bin Lou
    • 1
  • Fei Peng
    • 1
  • Aizhen Yang
    • 4
  • Xiaoning Wang
    • 1
  • Yicheng Ni
    • 1
    • 5
  • Jian Zhang
    • 1
  1. 1.Laboratory of Translational MedicineJiangsu Province Academy of Traditional Chinese MedicineNanjingChina
  2. 2.Department of OrthopaedicsZhongda Hospital, Southeast UniversityNanjingChina
  3. 3.Department of OrthopedicsThe Third Affiliated Hospital of Soochow University (The First People’s Hospital of Changzhou)Changzhou CityChina
  4. 4.Center Laboratory of Cancer CenterThe Jingdu Hospital of NanjingNanjingChina
  5. 5.Faculty of MedicineKU LeuvenLeuvenBelgium

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