Molecular Medicine

, Volume 19, Issue 1, pp 377–386 | Cite as

Antiangiogenic and Antitumoral Effects Mediated by a Vascular Endothelial Growth Factor Receptor 1 (VEGFR-1)-Targeted DNAzyme

  • Liangfang Shen
  • Qin Zhou
  • Ying Wang
  • Weihua Liao
  • Yan Chen
  • Zhijie Xu
  • Lifang Yang
  • Lun-Quan Sun
Research Article


Antiangiogenesis is a promising antitumor strategy that inhibits tumor vascular formation to suppress tumor growth. DNAzymes are synthetic single-strand deoxyribonucleic acid (DNA) molecules that can cleave ribonucleic acids (RNAs). Here, we conducted a comprehensive in vitroselection of active DNAzymes for their activity to cleave the vascular endothelial growth factor receptor (VEGFR-1) mRNA and screened for their biological activity in a matrigel tube-formation assay. Among the selected DNAzymes, DT18 was defined as a lead molecule that was further investigated in several model systems. In a rat corneal vascularization model, DT18 demonstrated significant and specific antiangiogenic activity, as evidenced by the reduced area and vessel number in VEGF-induced corneal angiogenesis. In a mouse melanoma model, DT18 was shown to inhibit B16 tumor growth, whereas it did not affect B16 cell proliferation. We further assessed the DT18 effect in mice with established human nasopharyngeal carcinoma (NPC). A significant inhibition of tumor growth was observed, which accompanied downregulation of VEGFR-1 expression in NPC tumor tissues. To evaluate DT18 effect on vasculature, we performed dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) on the human NPC xenograft mice treated with DT18 and showed a reduction of the parameter of Ktrans (volume constant for transfer of contrast agent), which reflects the condition of tumor microvascular permeability. When examining the safety and tolerability of DT18, intravenous administration of Dz18 to healthy mice caused no substantial toxicities, as shown by parameters such as body weight, liver/kidney function, and histological and biochemical analyses. Taken together, our data suggest that the anti-VEGFR-1DNAzyme may be used as a therapeutic agent for the treatment of cancer, such as NPC.



We would like to thank Crispin Dass for technical assistance in animal experimentation. This work was partly supported by the National Natural Science Foundation of China (81172188),91129709,81072220 and 81000596) and the PhD Graduate Supervision Fund of Ministry of Education, China (20110162110010).


  1. 1.
    Chung AS, Ferrara N. (2011) Developmental and pathological angiogenesis. Annual Review of Cell and Developmental Biology. 27:563–84.CrossRefPubMedGoogle Scholar
  2. 2.
    Adams RH, Alitalo K. (2007) Molecular regulation of angiogenesis and lymphangiogenesis. Nat. Rev. Mol. Cell Biol. 8:464–78.CrossRefPubMedGoogle Scholar
  3. 3.
    Augustin HG, Koh GY, Thurston G, Alitalo K. (2009) Control of vascular morphogenesis and homeostasis through the angiopoietin-Tie system. Nat. Rev. Mol. Cell Biol. 10:165–77.CrossRefPubMedGoogle Scholar
  4. 4.
    Potente M, Gerhardt H, Carmeliet P. (2011) Basic and therapeutic aspects of angiogenesis. Cell. 146:873–87.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Carmeliet P, Jain RK. (2011) Molecular mechanisms and clinical applications of angiogenesis. Nature. 473:298–307.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Friesel RE, Maciag T. (1995) Molecular mechanisms of angiogenesis: fibroblast growth factor signal transduction. FASEB J. 9:919–25.CrossRefPubMedGoogle Scholar
  7. 7.
    Gerber HP, Condorelli F, Park J, Ferrara N. (1997) Differential transcriptional regulation of the two vascular endothelial growth factor receptor genes: Flt-1, but not Flk-1/KDR, is up-regulated by hypoxia. J. Biol. Chem. 272:23659–67.CrossRefPubMedGoogle Scholar
  8. 8.
    Carmeliet P, et al. (2001) Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions. Nat. Med. 7:575–83.CrossRefPubMedGoogle Scholar
  9. 9.
    Cherrington JM, Strawn LM, Shawver LK. (2000) New paradigms for the treatment of cancer: the role of anti-angiogenesis agents. Adv. Cancer Res. 79:1–38.CrossRefPubMedGoogle Scholar
  10. 10.
    Ferrara N. (1997) The Role of Vascular Endothelial Growth Factor in the Regulation of Blood Vessel Growth. In: Tumour Angiogenesis. Bicknell R, Lewis CE, Ferrara N (eds.) Oxford University Press, New York, pp. 185–98.Google Scholar
  11. 11.
    Ellis LM, Takahashi Y, Liu W, Shaheen RM. (2000) Vascular endothelial growth factor in human colon cancer: biology and therapeutic implications. Oncologist. 5 Suppl 1:11–15.CrossRefPubMedGoogle Scholar
  12. 12.
    Ivy SP, Wick JY, Kaufman BM. (2009) An overview of small-molecule inhibitors of VEGFR signaling. Nat. Rev. Clin. Oncol. 6:569–79.CrossRefPubMedGoogle Scholar
  13. 13.
    Takahashi Y, Kitadai Y, Bucana CD, Cleary KR, Ellis LM. (1995) Expression of vascular endothelial growth factor and its receptor, KDR, correlates with vascularity, metastasis, and proliferation of human colon cancer. Cancer Res. 55:3964–8.PubMedGoogle Scholar
  14. 14.
    Chen CT, Hung MC. (2013) Beyond anti-VEGF: dual-targeting antiangiogenic and antiproliferative therapy. Am. J. Transl. Res. 5:393–403.PubMedPubMedCentralGoogle Scholar
  15. 15.
    Santoro SW, Joyce GF. (1997) A general purpose RNA-cleaving DNA enzyme. Proc. Natl. Acad. Sci. U. S. A. 94:4262–6.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Sun LQ, Cairns MJ, Saravolac EG, Baker A, Gerlach WL. (2000) Catalytic nucleic acids: from lab to applications. Pharmacol. Rev. 52:325–47.PubMedGoogle Scholar
  17. 17.
    Rossi JJ. (2012) Resurrecting DNAzymes as sequence-specific therapeutics. Sci. Transi. Med. 4:139fs120.Google Scholar
  18. 18.
    Bhindi R, et al. (2007) Brothers in arms: DNA enzymes, short interfering RNA, and the emerging wave of small-molecule nucleic acid-based genesilencing strategies. Am. J. Pathol. 171:1079–88.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Cairns MJ, Hopkins TM, Witherington C, Wang L, Sun LQ. (1999) Target site selection for an RNA-cleaving catalytic DNA. Nat. Biotechnol. 17:480–6.CrossRefPubMedGoogle Scholar
  20. 20.
    Santiago FS, et al. (1999) New DNA enzyme targeting Egr-1 mRNA inhibits vascular smooth muscle proliferation and regrowth after injury. Nat. Med. 5:1264–9.CrossRefPubMedGoogle Scholar
  21. 21.
    Yang L, Lu Z, Ma X, Cao Y, Sun LQ. (2010) A therapeutic approach to nasopharyngeal carcinomas by DNAzymes targeting EBV LMP-1 gene. Molecules. 15:6127–39.CrossRefPubMedGoogle Scholar
  22. 22.
    Cai H, et al. (2012) DNAzyme targeting c-jun suppresses skin cancer growth. Sci. Transl. Med. 4:139ra182.CrossRefGoogle Scholar
  23. 23.
    Cho EA, et al. (2013) Safety and tolerability of an intratumorally injected DNAzyme, Dz13, in patients with nodular basal-cell carcinoma: a phase 1 firstin-human trial (DISCOVER). Lancet. 381:1835–43.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Glaser R, et al. (1989) Two epithelial tumor cell lines (HNE-1 and HONE-1) latently infected with Epstein-Barr virus that were derived from nasopharyngeal carcinomas. Proc. Natl. Acad. Sci. U. S. A. 86:9524–8.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Santoro SW, Joyce GF. (1998) Mechanism and utility of an RNA-cleaving DNA enzyme. Biochemistry. 37:13330–42.CrossRefPubMedGoogle Scholar
  26. 26.
    Cairns MJ, King A, Sun LQ. (2003) Optimisation of the 10–23 DNAzyme-substrate pairing interactions enhanced RNA cleavage activity at purinecytosine target sites. Nucleic Acids Res. 31:2883–9.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Parry TJ, et al. (1999) Bioactivity of anti-angiogenic ribozymes targeting Flt-1 and KDR mRNA. Nucleic Acids Res. 27:2569–77.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Benimetskaya L, et al. (1998) Cationic porphyrins: novel delivery vehicles for antisense oligodeoxynucleotides. Nucleic Acids Res. 26:5310–5317.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Cheng HL. (2007) Dynamic contrast-enhanced MRI in oncology drug development. Curr. Clin. Pharmacol. 2:111–22.CrossRefPubMedGoogle Scholar
  30. 30.
    Agrawal S, et al. (1991) Pharmacokinetics biodistribution and stability of oligonucleotide phosphorothioates in mice. Proc. Natl. Acad. Sci. U. S. A. 88:7595–9.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Bellou S, Pentheroudakis G, Murphy C, Fotsis T. (2013) Anti-angiogenesis in cancer therapy: Hercules and hydra. Cancer Lett. 2:219–28.CrossRefGoogle Scholar
  32. 32.
    Ratner M. (2004) Genentech discloses safety concerns over Avastin. Nat. Biotechnol. 22:1198.CrossRefPubMedGoogle Scholar
  33. 33.
    Rahimi N. (2006) VEGFR-1 and VEGFR-2: two non-identical twins with a unique physiognomy. Front Biosci. 11:818–29.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Zhang L, et al. (2002) Angiogenic inhibition mediated by a DNAzyme that targets vascular endothelial growth factor receptor 2. Cancer Res. 62:5463–9.PubMedGoogle Scholar

Copyright information

© The Author(s) 2013

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (

Authors and Affiliations

  • Liangfang Shen
    • 1
  • Qin Zhou
    • 1
  • Ying Wang
    • 1
  • Weihua Liao
    • 2
  • Yan Chen
    • 4
  • Zhijie Xu
    • 3
  • Lifang Yang
    • 3
    • 4
  • Lun-Quan Sun
    • 4
  1. 1.Department of Oncology, Xiangya HospitalCentral South UniversityChangsha, HunanChina
  2. 2.Department of Radiology, Xiangya HospitalCentral South UniversityChangsha, HunanChina
  3. 3.Cancer Research InstituteCentral South UniversityChangsha, HunanChina
  4. 4.Center for Molecular Medicine, Xiangya HospitalCentral South UniversityChangsha, HunanChina

Personalised recommendations