Tumor Biology

, Volume 37, Issue 4, pp 4679–4688 | Cite as

Endostatin combined with radiotherapy suppresses vasculogenic mimicry formation through inhibition of epithelial–mesenchymal transition in esophageal cancer

  • Xiaochen Chen
  • Hao Zhang
  • Hongcheng Zhu
  • Xi Yang
  • Yuehua Yang
  • Yan Yang
  • Hua Min
  • Guangzong Chen
  • Jia Liu
  • Jing Lu
  • Hongyan Cheng
  • Xinchen SunEmail author
Original Article


The growth of solid tumors requires angiogenesis to provide oxygen and nutrients and to support cell proliferation. The switch from an avascular to a vascular phenotype is typically related to acceleration of tumor growth. Anti-angiogenic therapy is becoming a very promising way for malignant tumors. Meanwhile, malignant tumor cells themselves were able to develop the formation of cell-lined vessels that contribute to tumor neovascularization and supply the nutrients and oxygen, which is called vasculogenic mimicry (VM). However, the molecular mechanism of VM remains unclear. The purpose of this study was to investigate the efficacy of the novel recombinant human endostatin (rh-Endo) protein combined with radiotherapy on human esophageal squamous cell carcinoma (ESCC) cell lines Eca-109 and TE13. Our results showed that rh-Endo combined with radiotherapy significantly inhibited the proliferation, migration, invasion, and VM of human esophageal cancer cells in a dose-dependent manner; however, it has no direct effect on apoptosis of carcinoma cells, which indicated that rh-Endo combined with radiotherapy significantly changed the microenvironment of esophageal carcinoma, and played an important role in preventing distant metastasis. Our findings suggested that rh-Endo inhibited the metastasis of esophageal cancer and the activation of AKT pathway, and the down-regulation of epithelial–mesenchymal transition (EMT) may be associated with such effect of rh-Endo. These results also supported the bright prospect of rh-Endo combined with radiotherapy for clinical applications in the future.


Recombinant human endostatin Vasculogenic mimicry Radiotherapy Esophageal carcinoma Epithelial–mesenchymal transition 



Esophageal squamous cell carcinoma




Recombinant human endostatin


Non-small-cell lung cancer


Epithelial–mesenchymal transition


Vaculogenic mimicry


Dimethyl sulfoxide


Dulbecco’s modified eagle medium


Fluorescein isothiocyanate


Cell counting kit-8


Phosphatase and tensin homolog deleted on chromosome ten


Protein kinase B PKB


Glycogen synthase kinase 3β


Glyceraldehyde-3-phosphate dehydrogenase



This work was supported by the Natural Science Foundation of China (No. 81272504, No. 81472809), Innovation Team [No. LJ201123 (EH11)], A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) (JX10231801), grants from Key Academic Discipline of Jiangsu Province “Medical Aspects of Specific Environments”, Research and Innovation Project for College Graduates of Jiangsu Province (KYLX_0955). The methods were carried out in accordance with the approved guidelines.

Compliance with Ethical Standards

Conflicts of interest

The authors declare that they have no conflicts of interest.


  1. 1.
    Yu S, Yang CS, Li J, You W, Chen J, Cao Y, et al. Cancer prevention research in china. Cancer Prev Res (Phila). 2015;8:662–74.CrossRefGoogle Scholar
  2. 2.
    D’Journo XB, Thomas PA. Current management of esophageal cancer. J Thorac Dis. 2014;6 Suppl 2:S253–64.PubMedPubMedCentralGoogle Scholar
  3. 3.
    Jain RK. Normalizing tumor vasculature with anti-angiogenic therapy: a new paradigm for combination therapy. Nat Med. 2001;7:987–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Carmeliet P, Jain RK. Angiogenesis in cancer and other diseases. Nature. 2000;407:249–57.CrossRefPubMedGoogle Scholar
  5. 5.
    Behl T, Kaur I, Goel H, Kotwani A. Significance of the antiangiogenic mechanisms of thalidomide in the therapy of diabetic retinopathy. Vascul Pharmacol 2015.Google Scholar
  6. 6.
    Salza R, Oudart JB, Ramont L, Maquart FX, Bakchine S, Thoannes H, et al. Endostatin level in cerebrospinal fluid of patients with Alzheimer’s disease. J Alzheimers Dis. 2015;44:1253–61.PubMedGoogle Scholar
  7. 7.
    Al-Rawi S, Meehan-Andrews T, Bradley C, Al-Rawi J. Novel benzoxazines as inhibitors of angiogenesis. Investig New Drugs. 2015;33:45–52.CrossRefGoogle Scholar
  8. 8.
    Xu X, Mao W, Chen Q, Zhuang Q, Wang L, Dai J, et al. Endostar, a modified recombinant human endostatin, suppresses angiogenesis through inhibition of Wnt/beta-catenin signaling pathway. PLoS One. 2014;9:e107463.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Sun T, Zhao N, Zhao XL, Gu Q, Zhang SW, Che N, et al. Expression and functional significance of Twist1 in hepatocellular carcinoma: its role in vasculogenic mimicry. Hepatology. 2010;51:545–56.CrossRefPubMedGoogle Scholar
  10. 10.
    Kong D, Li Y, Wang Z, Sarkar FH. Cancer stem cells and epithelial-to-mesenchymal transition (EMT)-phenotypic cells: are they cousins or twins? Cancers (Basel). 2011;3:716–29.CrossRefGoogle Scholar
  11. 11.
    Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Invest. 2009;119:1420–8.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Polyak K, Weinberg RA. Transitions between epithelial and mesenchymal states: acquisition of malignant and stem cell traits. Nat Rev Cancer. 2009;9:265–73.CrossRefPubMedGoogle Scholar
  13. 13.
    Hassan H, Greve B, Pavao MS, Kiesel L, Ibrahim SA, Gotte M. Syndecan-1 modulates beta-integrin-dependent and interleukin-6-dependent functions in breast cancer cell adhesion, migration, and resistance to irradiation. FEBS J. 2013;280:2216–27.CrossRefPubMedGoogle Scholar
  14. 14.
    Meng MB, Jiang XD, Deng L, Na FF, He JZ, Xue JX, et al. Enhanced radioresponse with a novel recombinant human endostatin protein via tumor vasculature remodeling: experimental and clinical evidence. Radiother Oncol. 2013;106:130–7.CrossRefPubMedGoogle Scholar
  15. 15.
    He E, Pan F, Li G, Li J. Fractionated ionizing radiation promotes epithelial-mesenchymal transition in human esophageal cancer cells through PTEN deficiency-mediated akt activation. PLoS One. 2015;10:e126149.Google Scholar
  16. 16.
    Ricci-Vitiani L, Pallini R, Biffoni M, Todaro M, Invernici G, Cenci T, et al. Tumour vascularization via endothelial differentiation of glioblastoma stem-like cells. Nature. 2010;468:824–8.CrossRefPubMedGoogle Scholar
  17. 17.
    Dejana E. The role of wnt signaling in physiological and pathological angiogenesis. Circ Res. 2010;107:943–52.CrossRefPubMedGoogle Scholar
  18. 18.
    Otrock ZK, Mahfouz RA, Makarem JA, Shamseddine AI. Understanding the biology of angiogenesis: review of the most important molecular mechanisms. Blood Cells Mol Dis. 2007;39:212–20.CrossRefPubMedGoogle Scholar
  19. 19.
    Jung IL, Kang HJ, Kim KC, Kim IG. PTEN/pAkt/p53 signaling pathway correlates with the radioresponse of non-small cell lung cancer. Int J Mol Med. 2010;25:517–23.PubMedGoogle Scholar
  20. 20.
    Bai X, Li X, Tian J, Zhou Z. Antiangiogenic treatment diminishes renal injury and dysfunction via regulation of local AKT in early experimental diabetes. PLoS One. 2014;9:e96117.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Kim ES, Herbst RS. Angiogenesis inhibitors in lung cancer. Curr Oncol Rep. 2002;4:325–33.CrossRefPubMedGoogle Scholar
  22. 22.
    Cui C, Mao L, Chi Z, Si L, Sheng X, Kong Y, et al. A phase II, randomized, double-blind, placebo-controlled multicenter trial of Endostar in patients with metastatic melanoma. Mol Ther. 2013;21:1456–63.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Wang R, Chadalavada K, Wilshire J, Kowalik U, Hovinga KE, Geber A, et al. Glioblastoma stem-like cells give rise to tumour endothelium. Nature. 2010;468:829–33.CrossRefPubMedGoogle Scholar
  24. 24.
    Maniotis AJ, Folberg R, Hess A, Seftor EA, Gardner LM, Pe’Er J, et al. Vascular channel formation by human melanoma cells in vivo and in vitro: vasculogenic mimicry. Am J Pathol. 1999;155:739–52.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Du J, Sun B, Zhao X, Gu Q, Dong X, Mo J, et al. Hypoxia promotes vasculogenic mimicry formation by inducing epithelial-mesenchymal transition in ovarian carcinoma. Gynecol Oncol. 2014;133:575–83.CrossRefPubMedGoogle Scholar
  26. 26.
    Welch-Reardon KM, Wu N, Hughes CC. A role for partial endothelial-mesenchymal transitions in angiogenesis? Arterioscler Thromb Vasc Biol. 2015;35:303–8.CrossRefPubMedGoogle Scholar
  27. 27.
    Fan F, Samuel S, Evans KW, Lu J, Xia L, Zhou Y, et al. Overexpression of snail induces epithelial-mesenchymal transition and a cancer stem cell-like phenotype in human colorectal cancer cells. Cancer Med. 2012;1:5–16.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Keniry M, Parsons R. The role of PTEN signaling perturbations in cancer and in targeted therapy. Oncogene. 2008;27:5477–85.CrossRefPubMedGoogle Scholar
  29. 29.
    Kim J, Kang HS, Lee YJ, Lee HJ, Yun J, Shin JH, et al. EGR1-dependent PTEN upregulation by 2-benzoyloxycinnamaldehyde attenuates cell invasion and EMT in colon cancer. Cancer Lett. 2014;349:35–44.CrossRefPubMedGoogle Scholar
  30. 30.
    Roma-Rodrigues C, Fernandes AR, Baptista PV. Exosome in tumour microenvironment: Overview of the crosstalk between normal and cancer cells. Biomed Res Int. 2014;2014:179486.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Tickner JA, Urquhart AJ, Stephenson SA, Richard DJ, O’Byrne KJ. Functions and therapeutic roles of exosomes in cancer. Front Oncol. 2014;4:127.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Greening DW, Gopal SK, Mathias RA, Liu L, Sheng J, Zhu HJ, et al. Emerging roles of exosomes during epithelial-mesenchymal transition and cancer progression. Semin Cell Dev Biol. 2015;40:60–71.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Xiaochen Chen
    • 1
  • Hao Zhang
    • 1
  • Hongcheng Zhu
    • 1
  • Xi Yang
    • 1
  • Yuehua Yang
    • 1
  • Yan Yang
    • 1
  • Hua Min
    • 1
  • Guangzong Chen
    • 1
  • Jia Liu
    • 1
  • Jing Lu
    • 1
  • Hongyan Cheng
    • 2
  • Xinchen Sun
    • 1
    Email author
  1. 1.Department of Radiation OncologyThe First Affiliated Hospital of Nanjing Medical UniversityNanjingChina
  2. 2.Department of General Internal MedicineThe First Affiliated Hospital of Nanjing Medical UniversityNanjingChina

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