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A novel fusion suicide gene yeast CDglyTK plays a role in radio-gene therapy of nasopharyngeal carcinoma

Cancer Gene Therapy volume 11, pages 790–796 (2004)Cite this article

Abstract

To investigate a novel suicide gene for nasopharyngeal carcinoma (NPC) therapy, the yCDglyTK gene was constructed by fusing yeast cytosine deaminase (CD) and herpes simplex type 1 thymidine kinase. The expression of the yCDglyTK gene was detected by RT-PCR and Western blotting, and its bioactivity was demonstrated by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. An animal study was carried out in which BALB/C nude mice bearing yCDglyTK gene-modified tumors were treated with prodrugs and radiation. Our results revealed that the yCDglyTK gene could be expressed in CNE-2 cells in vitro. In MTT analysis, at the transfection rate of 10%, 66% cells were killed. The synergistic effect of CD and TK showed 91% of yCDglyTK-transfected cells were killed with the treatment of 5-fluorocytosine (5-FC) alone, 60% killed with ganciclovir (GCV) alone, and 75% killed with 5-FC and GCV together. In vivo, the tumor volume in all of the four prodrugs and/or radiation-treated groups were significantly different from that in the PBS-controlled group (P<.01); also yCDglyTK+prodrug+radiation group was different from the other three groups (P<.05). Our findings suggested there was a synergistic antitumor effect when combining suicide gene therapy and radiation, and yCDglyTK has potent antitumor efficacy and may be a candidate suicide gene for cancer therapy.

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Abbreviations

NPC:

nasopharyngeal carcinoma

CD:

cytosine deaminase

HSV-1 TK:

herpes simplex type 1 thymidine kinase

5-FC:

5-fluorocytosine

GCV:

ganciclovir

5-FdUMP:

5-fluoro-2′-deoxyuridine 5′-monophosphate

5-FUTP:

5-fluorouridine 5′-triphosphate

5-FUDP:

5-fluorouridine 5′-diphosphate

5-FdUDP:

5-fluoro-2′-deoxyuridine 5′-diphosphate

5-FdUTP:

5-fluoro-2′-deoxyuridine 5′-triphosphate

Egr-1:

early growth response factor 1

DPD:

dihydropyrimidine dehydrogenase

UPRT:

uracil phosphoribosyltransferase

References

  1. Hirayama T . Descriptive and analytical epidemiology of nasopharyngeal cancer. IARC Sci Publ. 1978;20:167–189.

    Google Scholar 

  2. Li CC, Yu MC, Henderson BE . Some epidemiologic observations of nasopharyngeal carcinoma in Guangdong, People's Republic of China. Natl Cancer Inst Monogr. 1985;69:49–52.

    CAS  PubMed  Google Scholar 

  3. Freytag SO, Khil M, Stricker H, et al. Phase I study of replication-competent adenovirus-mediated double suicide gene therapy for the treatment of locally recurrent prostate cancer. Cancer Res. 2002;62:4968–4976.

    CAS  PubMed  Google Scholar 

  4. Sung MW, Yeh HC, Thung SN, et al. Intratumoral adenovirus-mediated suicide gene transfer for hepatic metastases from colorectal adenocarcinoma: results of a phase I clinical trial. Mol Ther. 2001;4:182–191.

    Article  CAS  PubMed  Google Scholar 

  5. Klatzmann D, Cherin P, Bensimon G, et al. A phase I/II dose-escalation study of herpes simplex virus type 1 thymidine kinase “suicide” gene therapy for metastatic melanoma. Study Group on Gene Therapy of Metastatic Melanoma. Hum Gene Ther. 1998;9:2585–2594.

    CAS  PubMed  Google Scholar 

  6. 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 

  7. Smalley SR, Kimler BF, Evans RG . 5-Fluorouracil modulation of radiosensitivity in cultured human carcinoma cells. Int J Radiat Oncol Biol Phys. 1991;20:207–211.

    Article  CAS  PubMed  Google Scholar 

  8. Lawrence TS, Blackstock AW, McGinn C . The mechanism of action of radiosensitization of conventional chemotherapeutic agents. Semin Radiat Oncol. 2003;13:13–21.

    Article  PubMed  Google Scholar 

  9. Ueno M, Koyama F, Yamada Y, et al. Tumor-specific chemo–radio-gene therapy for colorectal cancer cells using adenovirus vector expressing the cytosine deaminase gene. Anticancer Res. 2001;21:2601–2608.

    CAS  PubMed  Google Scholar 

  10. Kanazawa T, Mizukami H, Okada T, et al. Suicide gene therapy using AAV-HSVtk/ganciclovir in combination with irradiation results in regression of human head and neck cancer xenografts in nude mice. Gene Therapy. 2003;10:51–58.

    Article  CAS  PubMed  Google Scholar 

  11. Joki T, Nakamura M, Ohno T . Activation of the radiosensitive EGR-1 promoter induces expression of the herpes simplex virus thymidine kinase gene and sensitivity of human glioma cells to ganciclovir. Hum Gene Ther. 1995;6:1507–1513.

    Article  CAS  PubMed  Google Scholar 

  12. Sukhatme VP, Cao XM, Chang LC, et al. A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization. Cell. 1988;53:37–43.

    Article  CAS  PubMed  Google Scholar 

  13. Datta R, Taneja N, Sukhatme VP, Qureshi SA, Weichselbaum R, Kufe DW . Reactive oxygen intermediates target CC(A/T)6GG sequences to mediate activation of the early growth response 1 transcription factor gene by ionizing radiation. Proc Natl Acad Sci USA. 1993;90:2419–2422.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Hallahan DE, Mauceri HJ, Seung LP, et al. Spatial and temporal control of gene therapy using ionizing radiation. Nat Med. 1995;1:786–791.

    Article  CAS  PubMed  Google Scholar 

  15. Kawashita Y, Ohtsuru A, Kaneda Y, et al. Regression of hepatocellular carcinoma in vitro and in vivo by radiosensitizing suicide gene therapy under the inducible and spatial control of radiation. Hum Gene Ther. 1999;10:1509–1519.

    Article  CAS  PubMed  Google Scholar 

  16. Greco O, Marples B, Dachs GU, Williams KJ, Patterson AV, Scott SD . Novel chimeric gene promoters responsive to hypoxia and ionizing radiation. Gene Therapy. 2002;9:1403–1411.

    Article  CAS  PubMed  Google Scholar 

  17. Li XJ, Wang KM, Xu Y, et al. Ionizing radiation-regulated killing of human hepatoma cells by liposome-mediated CDglyTK gene delivery. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003;35:64–70 (Chinese).

    CAS  Google Scholar 

  18. Kievit E, Nyati MK, Ng E, et al. Yeast cytosine deaminase improves radiosensitization and bystander effect by 5-fluorocytosine of human colorectal cancer xenografts. Cancer Res. 2000;60:6649–6655.

    CAS  PubMed  Google Scholar 

  19. Hamstra DA, Rice DJ, Fahmy S, Ross BD, Rehemtulla A . Enzyme/prodrug therapy for head and neck cancer using a catalytically superior cytosine deaminase. Hum Gene Ther. 1999;10:1993–2003.

    Article  CAS  PubMed  Google Scholar 

  20. Sizhong Z, Xiukung G, Yi Z . Cytogenetic studies on an epithelial cell line derived from poorly differentiated nasopharyngeal carcinoma. Int J Cancer. 1983;31:587–590.

    Article  CAS  PubMed  Google Scholar 

  21. Teng ZP, Ooka T, Huang DP, Zeng Y . Detection of Epstein-Barr Virus DNA in well and poorly differentiated nasopharyngeal carcinoma cell lines. Virus Genes. 1996;13:53–60.

    Article  CAS  PubMed  Google Scholar 

  22. Cole SP . Rapid chemosensitivity testing of human lung tumor cells using the MTT assay. Cancer Chemother Pharmacol. 1986;17:259–263.

    Article  CAS  PubMed  Google Scholar 

  23. Tiraby M, Cazaux C, Baron M, Drocourt D, Reynes JP, Tiraby G . Concomitant expression of E. coli cytosine deaminase and uracil phosphoribosyltransferase improves the cytotoxicity of 5-fluorocytosine. FEMS Microbiol Lett. 1998;167:41–49.

    Article  CAS  PubMed  Google Scholar 

  24. Adachi Y, Tamiya T, Ichikawa T, et al. Experimental gene therapy for brain tumors using adenovirus-mediated transfer of cytosine deaminase gene and uracil phosphoribosyltransferase gene with 5-fluorocytosine. Hum Gene Ther. 2000;11:77–89.

    Article  CAS  PubMed  Google Scholar 

  25. Miyagi T, Koshida K, Hori O, et al. Gene therapy for prostate cancer using the cytosine deaminase/uracil phosphoribosyltransferase suicide system. J Gene Med. 2003;5:30–37.

    Article  CAS  PubMed  Google Scholar 

  26. Xia JH, Xia K, Liang DS, et al. Combination of suicide gene therapy and radiation enhances killing of nasopharyngeal carcinoma xenograph. J Radiat Res. 2004;45:281–289.

    Article  CAS  PubMed  Google Scholar 

  27. Hoganson DK, Batra RK, Olsen JC, Boucher RC . Comparison of the effects of three different toxin genes and their levels of expression on cell growth and bystander effect in lung adenocarcinoma. Cancer Res. 1996;56:1315–1323.

    CAS  PubMed  Google Scholar 

  28. Trinh QT, Austin EA, Murray DM, Knick VC, Huber BE . Enzyme/prodrug gene therapy: comparison of cytosine deaminase/5-fluorocytosine versus thymidine kinase/ganciclovir enzyme/prodrug systems in a human colorectal carcinoma cell line. Cancer Res. 1995;55:4808–4812.

    CAS  PubMed  Google Scholar 

  29. Rogulski KR, Zhang K, Kolozsvary A, Kim JH, Freytag SO . Pronounced antitumor effects and tumor radiosensitization of double suicide gene therapy. Clin Cancer Res. 1997;3:2081–2088.

    CAS  PubMed  Google Scholar 

  30. Corban-Wilhelm H, Hull WE, Becker G, Bauder-Wust U, Greulich D, Debus J . Cytosine deaminase and thymidine kinase gene therapy in a Dunning rat prostate tumour model: absence of bystander effects and characterisation of 5-fluorocytosine metabolism with 19F-NMR spectroscopy. Gene Therapy. 2002;9(23):1564–1575.

    Article  CAS  PubMed  Google Scholar 

  31. Moriuchi S, Wolfe D, Tamura M, et al. Double suicide gene therapy using a replication defective herpes simplex virus vector reveals reciprocal interference in a malignant glioma model. Gene Therapy. 2002;9:584–591.

    Article  CAS  PubMed  Google Scholar 

  32. Erbs P, Regulier E, Kintz J, et al. In vivo cancer gene therapy by adenovirus-mediated transfer of a bifunctional yeast cytosine deaminase/uracil phosphoribosyltransferase fusion gene. Cancer Res. 2000;60:3813–3822.

    CAS  PubMed  Google Scholar 

  33. Chen YX, Xue ZG, Zheng D, et al. Sodium Chloride modified silica nanoparticles as a non-viral vector with a high efficiency of DNA transfer into cells. Curr Gene Ther. 2003;3:273–279.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Professor Stephen E Christmas from University of Liverpool for his critical review of the manuscript. This work was financially supported by the grants of State 863 project (No. 2002AA227011 and 2001AA217181), 973 project, state natural science foundation program (No. 31830200) and the Ministry of Education of China (No. 00-01).

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Authors and Affiliations

  1. National Lab. of Medical Genetics of China, Central South University, Changsha, Hunan, People's Republic of China

    Kun Xia, Desheng Liang, Aifa Tang, Qian Pan, Zhigao Long, Heping Dai, Fang Cai, Lingqian Wu & Jiahui Xia

  2. E.N.T. Department, Xiangya Hospital, Central South University, Changsha, Hunan, People's Republic of China

    Yong Feng, Junyi Zhang & Suping Zhao

  3. Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, Hunan, People's Republic of China

    Zhuchu Chen

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Correspondence to Jiahui Xia.

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Xia, K., Liang, D., Tang, A. et al. A novel fusion suicide gene yeast CDglyTK plays a role in radio-gene therapy of nasopharyngeal carcinoma. Cancer Gene Ther 11, 790–796 (2004). https://doi.org/10.1038/sj.cgt.7700728

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