Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

A new expression plasmid in Bifidobacterium longum as a delivery system of endostatin for cancer gene therapy

Cancer Gene Therapy volume 14, pages 151–157 (2007)Cite this article

Abstract

To utilize Bifidobacterium longum (B. longum) as a safe and stable delivery system for endostatin in cancer gene therapy, we constructed pBV22210 vector combining a chloramphenicol-resistance gene (Cmr) from pBCSK(+) plasmid, a cryptic plasmid pMB1 from B. longum strain with pBV222. Endostatin was cloned directly downstream of an N terminal His6-tag sequence in the pBV22210, so that the endostatin protein expressed in B. longum could be purified with Ni-binding resin. The results indicated that the plasmid electroporated into B. longum was maintained stably in the absence of selective antibiotics and did not significantly affect biological characteristics of B. longum. In addition, the plasmid in B. longum showed a strong inhibitory effect on the growth of mouse solid liver tumor in vivo. These results suggested that this new plasmid may be a stable vector in B. longum for transporting anti-cancer genes in cancer gene therapy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 1

Similar content being viewed by others

References

  1. Kimura NT, Taniguchi S, Aoki K, Baba T . Selective localization and growth of Bifidobacterium bifidum in mouse tumors following intravenous administration. Cancer Res 1980; 40: 2061–2068.

    CAS  PubMed  Google Scholar 

  2. Missich R, Sgorbati B, LeBlanc DJ . Transformation of Bifidobacterium longum with pRM2, a constructed Escherichia coli-B.longum shuttle vector. Plasmid 1994; 32: 208–211.

    Article  CAS  Google Scholar 

  3. Rossi M, Brigidi P, Matteuzzi D . An efficient transformation system for Bifidobacterium spp. Lett Appl Microbiol 1997; 24: 33–36.

    Article  CAS  Google Scholar 

  4. Rossi M, Brigidi P, Matteuzzi D . Improved cloning vectors for Bifidobacterium spp. Lett Appl Microbiol 1998; 26: 101–104.

    Article  CAS  Google Scholar 

  5. Rossi M, Brigidi P, Gonzalez Vara y Rodriguez A, Matteuzzi D . Characterization of the plasmid pMB1 from Bifidobacterium longum and its use for shuttle vector construction. Res Microbiol 1996; 147: 133–143.

    Article  CAS  Google Scholar 

  6. Park MS, Shin DW, Lee KH, Ji GE . Sequence analysis of plasmid pKJ50 from Bifidobacterium longum. Microbiology 1999; 145: 585–592.

    Article  CAS  Google Scholar 

  7. Yazawa K, Fujimori M, Nakamura T, Sasaki T, Amano J, Kano Y et al. Bifidobacterium longum as a delivery system for gene therapy of chemically induced rat mammary tumors. Breast Cancer Res Treat 2001; 66: 165–170.

    Article  CAS  Google Scholar 

  8. Nakamura T, Sasaki T, Fujimori M, Yazawa K, Kano Y, Amano J et al. Cloned cytosine deaminase gene expression of Bifidobacterium longum and application to enzyme/pro-drug therapy of hypoxic solid tumors. Biosci Biotechnol Biochem 2002; 66: 2362–2366.

    Article  CAS  Google Scholar 

  9. Li X, Fu GF, Fan YR, Liu WH, Liu XJ, Wang JJ et al. Bifidobacterium adolescentis as a delivery system of endostatin for cancer gene therapy: selective inhibitor of angiogenesis and hypoxic tumor growth. Cancer Gene Ther 2003; 10: 105–111.

    Article  CAS  Google Scholar 

  10. Fu GF, Li X, Hou YY, Fan YR, Liu WH, Xu GX . Bifidobacterium longum as an oral delivery system of endostatin for gene therapy on solid liver cancer. Cancer Gene Ther 2005; 12: 133–140.

    Article  CAS  Google Scholar 

  11. Yi C, Huang Y, Guo ZY, Wang SR . Antitumor effect of cytosine deaminase/5-fluorocytosine suicide gene therapy system mediated by Bifidobacterium infantis on melanoma. Acta Pharmacol Sin 2005; 26: 629–634.

    Article  CAS  Google Scholar 

  12. Wei YQ, Wang QR, Zhao X, Yang L, Tian L, Lu Y et al. Immunotherapy of tumors with xenogeneic endothelial cells as a vaccine. Nat Med 2000; 6: 1160–1166.

    Article  CAS  Google Scholar 

  13. Wei YQ, Huang MJ, Yang L, Zhao X, Tian L, Lu Y et al. Immunogene therapy of tumors with vaccine based on Xenopus homologous vascular endothelial growth factor as a model antigen. Proc Natl Acad Sci 2001; 98: 11545–11550.

    Article  CAS  Google Scholar 

  14. Wang J, Chen C, Li B, Yu H, Zhao Y, Sun J et al. Antioxidative function and biodistribution of [Gd@C82(OH)22]n nanoparticles in tumor-bearing mice. Biochem Pharmacol 2006; 71: 872–881.

    Article  CAS  Google Scholar 

  15. Boehm T, Folkman J, Browder T, O'Reilly MS . Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 1997; 390: 404–407.

    Article  CAS  Google Scholar 

  16. Smeianov VV, Efimov BA, Korschunov VM, Steele JL . Characterization of plasmid p4M from Bifidobacterium pseudocatenulatum. GenBank 2002; Accession No AF359574.

  17. Kaufmann P, Pfefferkorn A, Teuber M, Meile L . Complete nucleotide sequence of the 2.1 kb plasmid pAP1 isolated from Bifidobacterium asteroides. GenBank 1997; Accession No Y11549.

  18. O'Riordan K, Fitzgerald GF . Molecular characterisation of a 5.75-kb cryptic plasmid from Bifidobacterium breve NCFB 2258 and determination of mode of replication. FEMS Microbiol Lett 1999; 174: 285–294.

    Article  CAS  Google Scholar 

  19. Park MS, Lee KH, Ji GE . Isolation and characterization of two plasmids from Bifidobacterium longum. Lett Appl Microbiol 1997; 25: 5–7.

    Article  CAS  Google Scholar 

  20. Tanaka K, Samura K, Kano Y . Structural and functional analysis of pTB6 from Bifidobacterium longum. Biosci Biotechnol Biochem 2005; 69: 422–455.

    Article  CAS  Google Scholar 

  21. Gonzalez Vara A, Rossi M, Altomare L, Eikmanns B, Matteuzzi D . Stability of recombinant plasmids on the continuous culture of Bifidobacterium animalis ATCC 27536. Biotechnol Bioeng 2003; 84: 145–150.

    Article  Google Scholar 

  22. Zhu L, Li W, Dong X . Species identification of genus Bifidobacterium based on partial HSP60 gene sequences and proposal of Bifidobacterium thermacidophilum subsp. porcinum subsp. nov. Int J Syst Evol Microbiol 2003; 53: 1619–1623.

    Article  CAS  Google Scholar 

  23. 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  Google Scholar 

  24. Tjin Tham Sjin RM, Satchi-Fainaro R, Birsner AE, Ramanujam VM, Folkman J, Javaherian K . A 27-amino-acid synthetic peptide corresponding to the NH2-terminal zinc-binding domain of endostatin is responsible for its antitumor activity. Cancer Res 2005; 65: 3656–3663.

    Article  Google Scholar 

  25. Rothbard JB, Garlington S, Lin Q, Kirschberg T, Kreider E, McGrane PL et al. Conjugation of arginine oligomers to cyclosporin A facilitates topical delivery and inhibition of inflammation. Nat Med 2000; 6: 1253–1257.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Grants BM 2002009 from the Natural Science Foundation of Jiangsu Province, China to GXX, and Grants 30370462 and the NSFC/RGC Joint Research Scheme 30318004 from the National Natural Science Foundation of China, RFDP Grant 20050284025 from the State Educational Ministry of China and Grant BK2006713 from the Natural Science Foundation of Jiangsu Province, China to JJW. We thank Prof. Zhi-Wei Wu (Center for Public Health Research, Nanjing University) for reviewing the manuscript.

Author information

Authors and Affiliations

  1. Department of Biological Science and Technology and State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Mailbox 426, Nanjing University, Nanjing, China

    Y-F Xu, L-P Zhu, B Hu, G-F Fu, H-Y Zhang & J-J Wang

  2. Jiangsu Key Laboratory for Bioresource Technology, School of Life Sciences, Nanjing Normal University, Nanjing, China

    J-J Wang

  3. Center for Public Health Research, Nanjing University, Nanjing, China

    G-X Xu

  4. Jiangsu Research Center for Gene Pharmaceutical Engineering and Technology, Nanjing, China

    G-X Xu

Authors
  1. Y-F Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L-P Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. B Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G-F Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H-Y Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J-J Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. G-X Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to J-J Wang or G-X Xu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xu, YF., Zhu, LP., Hu, B. et al. A new expression plasmid in Bifidobacterium longum as a delivery system of endostatin for cancer gene therapy. Cancer Gene Ther 14, 151–157 (2007). https://doi.org/10.1038/sj.cgt.7701003

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.cgt.7701003

Keywords

This article is cited by

Search

Advanced search

Quick links