Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
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Potent Anticancer Effects of Lentivirus Encoding a Drosophila Melanogaster Deoxyribonucleoside Kinase Mutant Combined with Brivudine

  • Zhang, Nian-Qu (Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University) ;
  • Zhao, Lei (Center of Experiment Technology and Medical Research, China Medical University) ;
  • Ma, Shuai (Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University) ;
  • Gu, Ming (Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University) ;
  • Zheng, Xin-Yu (Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University)
  • Published : 2012.05.30
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Abstract

Objective: Deoxyribonucleoside kinase of Drosophila melanogaster (Dm-dNK) mutants have been reported to exert suicide gene effects in combined gene/chemotherapy of cancer. Here, we aimed to further evaluate the capacity of the mutanted enzyme and its potential for inhibiting cancer cell growth. Methods: We altered the sequence of the last 10 amino acids of Dm-dNK to perform site-directed mutagenesis and constructed active site mutanted Dm-dNK (Dm-dNKmut), RT-PCR and western bloting studies were used to reveal the expression of lentivirus mediated Dm-dNKmut in a breast cancer cell line (Bcap37), a gastric cancer cell line (SGC7901) and a colorectal cancer cell line (CCL187). [3H]-labeled substrates were used for enzyme activity assays, cell cytotoxicity was assessed by MTT assays, cell proliferation using a hemocytometer and apoptosis induction by thenannexin-V-FITC labeled FACS method. In vivo, an animal study was set out in which BALB/C nude mice bearing tumors were treated with lentivirus mediated expression of Dm-dNKmut with the pyrimidine nucleoside analog brivudine (BVDU, (E)-5-(2-bromovinyl)-(2-deoxyuridine). Results: The Dm-dNKmut could be stably expressed in the cancer cell lines and retained its enzymatic activity. Moreover, the cells expressing Dm-dNKmut exhibited increased sensitivity in combination with BVDU, with induction of apoptosis in vitro and in vivo. Conclusion: These findings underlined the importance of BVDU phosphorylated by Dm-dNKmut in transduced cancer cells and the potential role of Dm-dNKmut as a suicide gene, thus providing the basis for future intensive research for cancer therapy.

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References

  1. Caruso M, Panis Y, Gagandeep S, et al (1993). Regression of established macroscopic live metastases after in situ transduction of a suicide gene. Proc Natl Acad Sci USA, 90, 7024-8. https://doi.org/10.1073/pnas.90.15.7024
  2. Eichholz A, Merchant S, Gaya AM (2010). Anti-angiogenesis therapies: their potential in cancer management. Onco Targets Ther, 3, 69-82.
  3. Gerber DE (2008). Targeted therapies: a new generation of cancer treatments. Am Fam Physician, 77, 311-9.
  4. Ito M, Suda Y, Harashima H, Kamiya H (2010). Cytotoxic effect of Drosophila deoxynucleoside kinase gene on replicating plasmid in HeLa cells. Biol Pharm Bull, 33, 1223-7. https://doi.org/10.1248/bpb.33.1223
  5. Johansson M, van Rompay AR, Degreve B, Balzarini J, Karlsson A (1999). Cloning and characterization of the multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster. J Biol Chem, 274, 23814-9. https://doi.org/10.1074/jbc.274.34.23814
  6. Kamizono J, Nagano S, Murofushi Y, et al (2005). Survivinresponsive conditionally replicating adenovirus exhibits cancer-specific and efficient viral replication. Cancer Res, 65, 5284-91. https://doi.org/10.1158/0008-5472.CAN-04-2657
  7. Kianmanesh AR, Perrin H, Panis Y, et al (1997). A "distant" bystander effect of suicide gene therapy: regression of nontransduced tumors together with a distant transduced tumor. Hum Gene Ther, 8, 1807-14. https://doi.org/10.1089/hum.1997.8.15-1807
  8. Knecht W, Mikkelsen NE, Clausen AR, et al (2009). Drosophila melanogaster deoxyribonucleoside kinase activates gemcitabine. Biochem Biophys Res Commun, 382, 430-3. https://doi.org/10.1016/j.bbrc.2009.03.041
  9. Knecht W, Sandrini MP, Johansson K, et al (2002). A few amino acid substitutions can convert deoxyribonucleoside kinase specificity from pyrimidines to purines. EMBO J, 21, 1873-80. https://doi.org/10.1093/emboj/21.7.1873
  10. Ma S, Qu W, Mao L, et al (2012). Antitumor effects of oncolytic adenovirus armed with Drosophila melanogaster deoxyribonucleoside kinase in colorectal cancer. Oncol Rep, 27, 1443-50.
  11. Ma S, Zhao L, Zhu Z, et al (2011). The multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster as a therapeutic suicide gene of breast cancer cells. J Gene Med, 13, 305-11.
  12. Munch-Petersen B, Knecht W, Lenz C, Sondergaard L, Piskur J (2000). Functional expression of a multisubstrate deoxyribonucleoside kinase from Drosophila melanogaster and its C-terminal deletion mutants. J Biol Chem, 275, 6673-9. https://doi.org/10.1074/jbc.275.9.6673
  13. Munch-Petersen B, Piskur J, Sondergaard L (1998a). Four deoxynucleoside kinase activities from Drosophila melanogaster are contained within a single monomeric enzyme, a new multifunctional deoxynucleoside kinase. J Biol Chem, 273, 3926-31. https://doi.org/10.1074/jbc.273.7.3926
  14. Munch-Petersen B, Piskur J, Sondergaard L (1998b). The single deoxynucleoside kinase in Drosophila melanogaster, Dm-dNK, is multifunctional and differs from the mammalian deoxynucleoside kinases. Adv Exp Med Biol, 431, 465-9. https://doi.org/10.1007/978-1-4615-5381-6_92
  15. Nettelbeck DM, Rivera AA, Balague C, Alemany R, Curiel DT (2002). Novel oncolytic adenoviruses targeted to melanoma: specific viral replication and cytolysis by expression of E1A mutants from the tyrosinase enhancer/ promoter. Cancer Res, 62, 4663-70.
  16. Okabe S, Arai T, Yamashita H, Sugihara K (2003). Adenovirus-mediated prodrug-enzyme therapy for CEAproducing colorectal cancer cells. J Cancer Res Clin Oncol, 129, 367-73. https://doi.org/10.1007/s00432-003-0444-8
  17. Sandrini MP, Clausen AR, On SL, et al (2007). Nucleoside analogues are activated by bacterial deoxyribonucleoside kinases in a species-specific manner. J Antimicrob Chemother, 60, 510-20. https://doi.org/10.1093/jac/dkm240
  18. Solaroli N, Bjerke M, Amiri MH, Johansson M, Karlsson A (2003). Active site mutants of Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. Eur J Biochem, 270, 2879-84. https://doi.org/10.1046/j.1432-1033.2003.03666.x
  19. Solaroli N, Johansson M, Balzarini J, Karlsson A (2007). Enhanced toxicity of purine nucleoside analogs in cells expressing Drosophila melanogaster nucleoside kinase mutants. Gene Therapy, 14, 86-92. https://doi.org/10.1038/sj.gt.3302835
  20. Springer CJ, Niculescu-Duvaz I (2000). Prodrug-activating systems in suicide gene therapy. JJ Clin Invest, 105, 1161-7. https://doi.org/10.1172/JCI10001
  21. Springett GM, Moen RC, Anderson S, Blaese RM, Anderson WF (1989). Infection efficiency of T lymphocytes with amphotropic retroviral vectors is cell cycle dependent. J Virol, 63, 3865-9.
  22. Zheng X, Johansson M, Karlsson A (2000). Retroviral transduction of cancer cell lines with the gene encoding Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. J Biol Chem, 275, 39125-9. https://doi.org/10.1074/jbc.M006212200
  23. Zheng X, Johansson M, Karlsson A (2001a). Bystander effects of cancer cell lines transduced with the multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster and synergistic enhancement by hydroxyurea. Mol Pharmacol, 60, 262-6. https://doi.org/10.1124/mol.60.2.262
  24. Zheng X, Johansson M, Karlsson A (2001b). Nucleoside analog cytotoxicity and bystander cell killing of cancer cells expressing Drosophila melanogaster deoxyribonucleoside kinase in the nucleus or cytosol. Biochem Biophys Res Commun, 289, 229-33. https://doi.org/10.1006/bbrc.2001.5953
  25. Zhu Z, Ma S, Zhao L, et al (2011a). Adenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase mutants combined with gemcitabine harbor a safe cancer treatment profile. Int J Oncol, 38, 745-53.
  26. Zhu Z, Mao L, Zhao L, et al (2011b). Synergistic therapeutic effect in gastric cancer cells produced by oncolytic adenovirus encoding Drosophila melanogaster deoxyribonucleoside kinase. Cancer Biol Ther, 11, 874-82. https://doi.org/10.4161/cbt.11.10.15182

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