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.

  • Acquired Diseases
  • Published:

Fractionated radiation therapy in combination with adenoviral delivery of the cytosine deaminase gene and 5-fluorocytosine enhances cytotoxic and antitumor effects in human colorectal and cholangiocarcinoma models

Gene Therapy volume 7, pages 1019–1026 (2000)Cite this article

Abstract

Radiosensitization of human gastrointestinal tumors by 5-fluorouracil (5-FU) has been studied in vitro and clinically in human cancer therapy trials. The bacterial enzyme cytosine deaminase (CD) converts the nontoxic prodrug 5-fluorocytosine (5-FC) into 5-FU. Human colon cancer cells stably expressing CD have been shown by other investigators to be sensitized to radiation following treatment with 5-FC. We previously used an adenoviral vector under control of the cytomegalovirus promoter (AdCMVCD) encoding the CD gene in combination with 5-FC and a single fraction of radiation exposure to enhance cytotoxicity to human cholangiocarcinoma cells in vitro and in vivo. The purpose of this study was to determine whether AdCMVCD infection and 5-FC with multiple fraction low-dose radiotherapy results in enhanced cytotoxicity. In the present study, we utilized AdCMVCD and 5-FC with single fraction radiotherapy to demonstrate enhanced cytotoxicity to WiDr human colon carcinoma cells in vitro. Additionally, we tested this gene therapy/prodrug treatment strategy employing a fractionated radiation dosing schema in animal models of WiDr colon carcinoma and SK-ChA-1 cholangiocarcinoma. A prolonged WiDr tumor regrowth delay was obtained with AdCMVCD infection in combination with systemic delivery of 5-FC and fractionated external beam radiation therapy compared with control animals treated without radiation, without 5-FC, or without AdCMVCD. The results of treatment with AdCMVCD + 5-FC + radiation therapy to cholangiocarcinoma xenografts were equivalent to those obtained with systemic 5-FU administration + radiation. Thus, the use of AdCMVCD can be effectively combined with clinically relevant 5-FC and radiation administration schemes to achieve enhanced tumor cell killing and increased control of established tumors of human gastrointestinal malignancies.

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 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Lawrence TS, Tepper JE . Combined radiation therapy and chemotherapy – 5-FU and FUdR Semin Radiat Oncol 1997 7: 247–334

    Article  CAS  PubMed  Google Scholar 

  2. Hsue V et al. A phase I study of combined radiation therapy with 5-fluorouracil and low dose folinic acid in patients with locally advanced pancreatic or biliary carcinoma Int J Radiat Oncol Biol Phys 1996 34: 445–450

    Article  CAS  PubMed  Google Scholar 

  3. Oberfield RA, Rossi RL . The role of chemotherapy in the treatment of bile duct cancer World J Surg 1988 12: 105–108

    Article  CAS  PubMed  Google Scholar 

  4. Buchsbaum DJ et al. Approaches to enhance cancer radiotherapy employing gene transfer methods Gene Therapy 1996 3: 1042–1068

    CAS  PubMed  Google Scholar 

  5. Weichselbaum RR et al. Gene therapy targeted by radiation preferentially radiosensitizes tumor cells Cancer Res 1994 54: 4266–4269

    CAS  PubMed  Google Scholar 

  6. Kim JH, Kim SH, Brown SL, Freytag SO . Selective enhancement by an antiviral agent of the radiation-induced cell killing of human glioma cells transduced with HSV-tk gene Cancer Res 1994 54: 6053–6056

    CAS  PubMed  Google Scholar 

  7. Khil MS et al. Radiosensitization by 5-fluorocytosine of human colorectal carcinoma cells in culture transduced with cytosine deaminease gene Clin Cancer Res 1996 2: 53–57

    CAS  PubMed  Google Scholar 

  8. Pederson LC et al. Molecular chemotherapy combined with radiation therapy enhances killing of cholangiocarcinoma cells in vitro and in vivo Cancer Res 1997 57: 4325–4332

    CAS  PubMed  Google Scholar 

  9. Spitz FR et al. Adenoviral-mediated wild-type p53 gene expression sensitizes colorectal cancer cells to ionizing radiation Clin Cancer Res 1996 2: 1665–1671

    CAS  PubMed  Google Scholar 

  10. Huber BE et al. In vivo antitumor activity of 5-fluorocytosine on human colorectal carcinoma cells genetically modified to express cytosine deaminase Cancer Res 1993 53: 4619–4626

    CAS  PubMed  Google Scholar 

  11. Pederson LC et al. Combining cytosine deaminase expression, 5-fluorocytosine exposure, and radiotherapy increases cytotoxicity to cholangiocarcinoma cells J Gastrointest Surg 1998 2: 283–291

    Article  CAS  PubMed  Google Scholar 

  12. Rogulski KR, Kim JH, Kim SH, Freytag SO . Glioma cells transduced with an Escherichia coli CD/HSV-1 TK fusion gene exhibit enhanced metabolic suicide and radiosensitivity Hum Gene Ther 1997 8: 73–85

    Article  CAS  PubMed  Google Scholar 

  13. Dong Y et al. In vivo replication-deficient adenovirus vector-mediated transduction of the cytosine deaminase gene sensitizes glioma cells to 5-fluorocytosine Hum Gene Ther 1996 7: 713–720

    Article  CAS  PubMed  Google Scholar 

  14. Hirschowitz EA et al. In vivo adenovirus-mediated gene transfer of the Escherichia coli cytosine deaminase gene to human colon carcinoma-derived tumors induces chemosensitivity to 5-fluorocytosine Hum Gene Ther 1995 6: 1055–1063

    Article  CAS  PubMed  Google Scholar 

  15. Ohwada A, Hirschowitz EA, Crystal RG . Regional delivery of an adenovirus vector containing the Escherichia coli cytosine deaminase gene to provide local activation of 5-fluorocytosine to suppress the growth of colon carcinoma metastatic to liver Hum Gene Ther 1996 7: 1567–1576

    Article  CAS  PubMed  Google Scholar 

  16. Saunders K et al. Diffuse bile duct tumors: guidelines for management Am Surg 1991 57: 816–820

    CAS  PubMed  Google Scholar 

  17. Vauthey J-N, Blumgart LH . Recent advances in the management of cholangiocarcinomas Semin Liver Dis 1994 14: 109–114

    Article  CAS  PubMed  Google Scholar 

  18. Denning C, Pitts JD . Bystander effects of different enzyme-prodrug systems for cancer gene therapy depend on differentpathways for intercellular transfer of toxic metabolites, a factor that will govern clinical choice of appropriate regimes Hum Gene Ther 1997 8: 1825–1835

    Article  CAS  PubMed  Google Scholar 

  19. Paillard F . Bystander effects in enzyme/prodrug gene therapy Hum Gene Ther 1997 8: 1733–1736

    Article  CAS  PubMed  Google Scholar 

  20. Becker TC et al. Use of recombinant adenovirus for metabolic engineering of mammalian cells Meth Cell Biol 1994 43: 161–189

    Article  CAS  Google Scholar 

  21. Grizzle WE, Myers RB, Manne U, Srivastava S . Immunohistochemical evaluation of biomarkers in prostatic and colorectal neoplasia. In: Hanausek M, Walaszek Z (eds) John Walker's Methods in Molecular Medicine – Tumor Marker Protocols Humana Press: Totowa, NJ 1998 143–160

    Google Scholar 

  22. Grizzle WE et al. Factors affecting immunohistochemical evaluation of biomarker expression in neoplasia. In: Hanausek M, Walaszek Z (eds) John Walker's Methods in Molecular Medicine – Tumor Marker Protocols Humana Press: Totowa 1998 161–179

    Google Scholar 

  23. Manne U et al. Prognostic significance of Bcl-2 expression and p53 nuclear accumulation in colorectal adenocarcinoma Int J Cancer 1997 74: 346–358

    Article  CAS  PubMed  Google Scholar 

  24. Myers RB, Grizzle WE . Biomarker expression in prostatic intraepithelial neoplasia Eur Urol 1996 30: 153–166

    Article  CAS  PubMed  Google Scholar 

  25. Haack K et al. Detection of cytosine deaminase in genetically modified tumor cells by specific antibodies Hum Gene Ther 1997 8: 1395–1401

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We acknowledge the efforts of Jonathan Harris, Helen Hurst and Craig Mullen for providing the CD plasmid construct pTH-CD500, which was used to construct AdCMVCD. We thank Sreekanth Kancharla, Sheila Bright, Christine Olsen, and Rob Stockard for excellent technical assistance, and Sally Lagan for manuscript preparation.

Author information

Authors and Affiliations

  1. Department of Radiation Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    MA Stackhouse & DJ Buchsbaum

  2. Department of Surgery, University of Alabama at Birmingham, Birmingham, AL, USA

    LC Pederson & SM Vickers

  3. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    WE Grizzle

  4. Department of Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL, USA

    DT Curiel

  5. Ruprecht-Karls-Universität Heidelberg Klinikum, Heidelberg, Germany

    J Gebert & K Haack

  6. Department of Preventive Medicine, University of Kansas Medical Center, Kansas City, KS, USA

    MS Mayo

Authors
  1. MA Stackhouse
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. LC Pederson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. WE Grizzle
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. DT Curiel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J Gebert
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K Haack
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. SM Vickers
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. MS Mayo
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. DJ Buchsbaum
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stackhouse, M., Pederson, L., Grizzle, W. et al. Fractionated radiation therapy in combination with adenoviral delivery of the cytosine deaminase gene and 5-fluorocytosine enhances cytotoxic and antitumor effects in human colorectal and cholangiocarcinoma models. Gene Ther 7, 1019–1026 (2000). https://doi.org/10.1038/sj.gt.3301196

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301196

Keywords

This article is cited by

Search

Advanced search

Quick links