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  • Original Article
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Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth

Cancer Gene Therapy volume 19, pages 110–117 (2012)Cite this article

Abstract

Conventional plasmids for gene therapy produce low-level and short-term gene expression. Here, we first created minicircle carrying endostatin (mc-hES) for measurement of transfection efficiency. Compared with pcDNA-hES, MC-mediated endostatin gene transfer in vitro resulted in seven-fold greater endostatin expression levels in transfected cells and inhibited the growth of Human umbilical vein endothelial cells (HUVEC) more efficiently. HUVEC cell migration and tube-formation assays suggested that MC-mediated endostatin gene has significant anti-migration and anti-tube-formation capacity than that in pcDNA-hES. In vivo experiments showed that after transfection, mc-hES inhibited the growth of nasopharyngeal carcinoma xenografts. The tumor inhibition rates of mc-hES and pcDNA-hES were 60.8% and 26.9%, respectively (P<0.05). MC-mediated intratumoral endostatin expression in vivo was 2.2–17.9 times higher than pcDNA-hES in xenografted mice and lasted for 20 days. Our results suggest that minicircle DNA vectors might be a promising vector for biotherapy and should be further investigated.

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References

  1. Qin DX, Hu YH, Yan JH, Xu GZ, Cai WM, Wu XL et al. Analysis of 1379 patients with nasopharyngeal carcinoma treated by radiation. Cancer 1998; 61: 1117–1124.

    Article  Google Scholar 

  2. Lee AW, Poon YF, Foo W, Law SC, Cheung FK, Chan DK et al. Retrospective analysis of 5037 patients with nasopharyngeal carcinoma treated during 1976–1985: overall survival and patterns of failure. Int J Radiat Oncol Biol Phys 1992; 23: 261–270.

    Article  CAS  PubMed  Google Scholar 

  3. Teo PM, Kwan WH, Lee WY, Leung SF, Johnson PJ . Prognosticators determining survival subsequent to distant metastasis from nasopharyngeal carcinoma. Cancer 1996; 77: 2423–2431.

    Article  CAS  PubMed  Google Scholar 

  4. Yang FO, Lan KL, Chen CT, Liu JC, Weng CL, Chou CK et al. Endostatin-cytosine deaminase fusion protein suppresses tumor growth by targeting neovascular endothelial cells. Cancer Res 2006; 66: 378–384.

    Article  Google Scholar 

  5. O'Reilly MS, Boehm T, Shing Y, Fukai N, Vasios G, Lane WS et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell 1997; 88: 277–285.

    Article  CAS  PubMed  Google Scholar 

  6. Huang X, Wong MK, Zhao Q, Zhu Z, Wang KZ, Huang N et al. Soluble recombinant endostatin purified from Escherichia coli: antiangiogenic activity and antitumor effect. Cancer Res 2001; 61: 478–481.

    CAS  PubMed  Google Scholar 

  7. Harms JF, Welch DR, Samant RS, Shevde LA, Miele ME, Babu GR et al. A small molecule antagonist of the α5β3 integrin suppresses MDA-MB-435 skeletal metastasis. Clin Exp Metastasis 2004; 21: 119–128.

    Article  CAS  PubMed  Google Scholar 

  8. Huang M, Chen ZY, Hu SJ, Jia FJ, Li ZJ, Hoyt G et al. Novel minicircle vector for gene therapy in murine myocardial infarction. Circulation 2009; 120: S230–S237.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Pannell D, Ellis J . Silencing of gene expression: implications for design of retrovirus vectors. Rev Med Virol 2001; 11: 205–217.

    Article  CAS  PubMed  Google Scholar 

  10. Wang L, Cao O, Swalm B, Dobrzynski E, Mingozzi F, Herzog RW . Major role of local immune responses in antibody formation to factor IX in AAV gene transfer. Gene Ther 2005; 12: 1453–1464.

    Article  CAS  PubMed  Google Scholar 

  11. Zaiss AK, Muruve DA . Immune responses to adeno-associated virus vectors. Curr Gene Ther 2005; 5: 323–331.

    Article  CAS  PubMed  Google Scholar 

  12. Hacein-Bey-Abina S, Kalle CV, Schmidt M, McCormack MP, Wulffraat N, Leboulch P et al. LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science 2003; 302: 415–419.

    CAS  PubMed  Google Scholar 

  13. Ohlfest JR, Demorest ZL, Motooka Y, Vengco I, Oh S, Chen E et al. Combinatorial antiangiogenic gene therapy by nonviral gene transfer using the sleeping beauty transposon causes tumor regression and improves survival in mice bearing intracranial human glioblastoma. Mol Ther 2005; 12: 778–788.

    Article  CAS  PubMed  Google Scholar 

  14. Bigger BW, Tolmachov O, Collombet JM, Fragkos M, Palaszewski I, Coutelle C . An araC-controlled bacterial cre expression system to produce DNA minicircle vectors for nuclear and mitochondrial gene therapy. J Biol Chem 2001; 276: 23018–23027.

    Article  CAS  PubMed  Google Scholar 

  15. Darquet AM, Cameron B, Wils P, Scherman D, Crouzet J . A new DNA vehicle for nonviral gene delivery: supercoiled minicircle. Gene Ther 1997; 4: 1341–1349.

    Article  CAS  PubMed  Google Scholar 

  16. Darquet AM, Rangara R, Kreiss P, Schwartz B, Naimi S, Delaère P et al. Minicircle: an improved DNA molecule for in vitro and in vivo gene transfer. Gene Ther 1999; 6: 209–218.

    Article  CAS  PubMed  Google Scholar 

  17. Tolmachov O, Palaszewski I, Bigger B, Coutelle C . RecET driven chromosomal gene targeting to generate a RecA deficient Escherichia coli strain for Cre mediated production of minicircle DNA. BMC Biotechnol 2006; 6: 17–28.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Chen ZY, He CY, Ehrhardt A, Kay MA . Minicircle DNA vectors devoid of bacterial DNA result in persistent and high-level transgene expression in vivo. Mol Ther 2003; 8: 495–500.

    Article  CAS  PubMed  Google Scholar 

  19. Chen ZY, He CY, Meuse L, Kay MA . Silencing of episomal transgene expression by plasmid bacterial DNA elements in vivo. Gene Ther 2004; 11: 856–864.

    Article  CAS  PubMed  Google Scholar 

  20. Chen ZY, He CY, Kay MA . Improved production and purification of minicircle DNA vector free of plasmid bacterial sequences and capable of persistent transgene expression in vivo. Hum Gene Ther 2005; 16: 126–131.

    Article  CAS  PubMed  Google Scholar 

  21. Wu JX, Xiao X, Zhao P, Xue G, Zhu YH, Zhu X et al. Minicircle-IFNγ induces antiproliferative and antitumoral effects in human nasopharyngeal carcinoma. Clin Cancer Res 2006; 12: 4702–4713.

    Article  CAS  PubMed  Google Scholar 

  22. Zhao P, Luo R, Wu J, Xie F, Li H, Xiao X et al. E10A, an adenovirus carrying human endostatin gene, in combination with docetaxel treatment inhibits prostate cancer growth and metastases. J Cell Mol Med 2010; 14: 381–391.

    Article  CAS  PubMed  Google Scholar 

  23. Edelstein ML, Abedi MR, Wixon J . Gene therapy clinical trials worldwide to 2007—an update. J Gene Med 2007; 9: 833–842.

    Article  PubMed  Google Scholar 

  24. Chen ZY, Riu E, He CY, Xu H, Kay MA . Silencing of episomal transgene expression in liver by plasmid bacterial backbone DNA is independent of CpG methylation. Mol Ther 2008; 16: 548–556.

    Article  CAS  PubMed  Google Scholar 

  25. Masataka S, Kazue K, Yoshinaga S . Plasmid DNA sequences present in conventional herpes simplex virus amplicon vectors cause rapid transgene silencing by forming inactive chromatin. J Virol 2006; 4: 3293–3300.

    Google Scholar 

  26. Citrin D, Lee AK, Scott T . In vivo tumor imaging in mice with near-infrared labeled endostatin. Mol Cancer Ther 2004; 3: 481–488.

    CAS  PubMed  Google Scholar 

  27. Wickstrom SA, Alitalo K, Keski JO . Endostatin associates with integrin α5β1 and caveolin-1, and activates Src via a tyrosyl phosphatase-dependent pathway in human endothelial cells. Cancer Res 2002; 62: 5580–5589.

    CAS  PubMed  Google Scholar 

  28. Rehn M, Veikkola T, Kukk EV . Interaction of endostatin with integrins implicated in angiogenesis. Proc Nat Acad Sci USA 2001; 98: 1024–1029.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We are most grateful to Dr Zhiying Chen (Stanford University, Stanford, CA), who generously provided p2ϕC31 and advice on this work; we are also grateful to Dr Saiwah Tsao (University of Hong Kong, PR China) who generously provided HUVEC. This work was supported by National Natural Science Foundation of China (Grant No. 81001212 and 30801360), National Basic Research Program of China (973 Program Grant No.2010CB529904 and 2010CB912201), Foundation of Zhejiang Provincial Educational Committee (Grant No. Y201019175) and Zhejiang Provincial Health Bureau Foundation (Grant No. 2010KYB036).

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Author notes

  1. B-L Xu, L Yuan and J-X Wu: These authors contributed equally to this work.

Authors and Affiliations

  1. The Affiliated Tumor Hospital, Zhengzhou University, Zhengzhou, PRC

    B-L Xu & L Yuan

  2. State Key Laboratory of Oncology in South China, Cancer Center, Sun Yat-sen University, Guangzhou, PRC

    B-L Xu, J-X Wu & W-L Huang

  3. Department of Medical Oncology, The First Affiliated Hospital, Zhejiang University, Hangzhou, PRC

    N Xu, W-J Fang & P Zhao

  4. Institute of Microbiology, Chinese-Academy of Science, Beijing, PRC

    W-L Huang

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  1. B-L Xu
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  2. L Yuan
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  6. P Zhao
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Correspondence to P Zhao or W-L Huang.

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Xu, BL., Yuan, L., Wu, JX. et al. Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth. Cancer Gene Ther 19, 110–117 (2012). https://doi.org/10.1038/cgt.2011.72

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