Combretastatin A4 phosphate treatment induces vasculogenic mimicry formation of W256 breast carcinoma tumor in vitro and in vivo
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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.
KeywordsVasculogenic 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
- 19.Wang H, Sun X, Chen F, De Keyzer F, Yu J, Landuyt W, et al. Treatment of rodent liver tumor with combretastatin a4 phosphate: noninvasive therapeutic evaluation using multiparametric magnetic resonance imaging in correlation with microangiography and histology. Invest Radiol. 2009;44(1):44–53. doi: 10.1097/RLI.0b013e31818e5ace.CrossRefPubMedGoogle Scholar
- 20.Li J, Chen F, Feng Y, Cona MM, Yu J, Verbruggen A, et al. Diverse responses to vascular disrupting agent combretastatin a4 phosphate: a comparative study in rats with hepatic and subcutaneous tumor allografts using MRI biomarkers, microangiography, and histopathology. Transl Oncol. 2013;6(1):42–50. doi: 10.1593/tlo.12367.CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Dowlati A, Robertson K, Cooney M, Petros WP, Stratford M, Jesberger J, et al. A phase I pharmacokinetic and translational study of the novel vascular targeting agent combretastatin a-4 phosphate on a single-dose intravenous schedule in patients with advanced cancer. Cancer Res. 2002;62(12):3408–16.PubMedGoogle Scholar
- 34.Ljungkvist AS, Bussink J, Rijken PF, Kaanders JH, van der Kogel AJ, Denekamp J. Vascular architecture, hypoxia, and proliferation in first-generation xenografts of human head-and-neck squamous cell carcinomas. Int J Radiat Oncol Biol Phys. 2002;54(1):215–28. doi: 10.1016/S0360-3016(02)02938-3.CrossRefPubMedGoogle Scholar
- 41.Shaked Y, Tang T, Woloszynek J, Daenen LG, Man S, Xu P, et al. Contribution of granulocyte colony-stimulating factor to the acute mobilization of endothelial precursor cells by vascular disrupting agents. Cancer Res. 2009;69(19):7524–8. doi: 10.1158/0008-5472.CAN-09-0381.CrossRefPubMedPubMedCentralGoogle Scholar
- 54.Seftor RE, Seftor EA, Koshikawa N, Meltzer PS, Gardner LM, Bilban M, et al. Cooperative interactions of laminin 5 γ2 chain, matrix metalloproteinase-2, and membrane type-1-matrix/metalloproteinase are required for mimicry of embryonic vasculogenesis by aggressive melanoma. Cancer Res. 2001;61(17):6322–7.PubMedGoogle Scholar