Conditionally replicating adenovirus SG500-expressed mutant Dm-dNK gene for breast cancer therapy

Qu,Wenzhi; Zhu,Zhi; Zhao,Lei; He,Anning; Zheng,Xinyu

doi:10.3892/ijo.2012.1657

December 2012 Volume 41 Issue 6

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December 2012 Volume 41 Issue 6

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Article

Conditionally replicating adenovirus SG500-expressed mutant Dm-dNK gene for breast cancer therapy

Authors:
- Wenzhi Qu
- Zhi Zhu
- Lei Zhao
- Anning He
- Xinyu Zheng
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Affiliations: Department of Breast Surgery, Fourth Affiliated Hospital, China Medical University, Shenyang, P.R. China, Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University, Shenyang, P.R. China, Center of Experiment Technology and Medical Research, China Medical University, Shenyang, P.R. China, Laboratory 1, Cancer Institute, China Medical University, Shenyang, P.R. China
Pages: 2175-2183
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Published online on: October 11, 2012

https://doi.org/10.3892/ijo.2012.1657
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Abstract

The purpose of this analysis was to investigate the enzyme activity and specificity of using adenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) mutants in combination with gemcitabine. Compared with herpes simplex type 1 thymidine kinase (HSV-TK) and other known dNKs, this Dm-dNK enzyme has a broader substrate specificity and a higher catalytic rate. We created the Dm-dNK mutants (dNKmu) by site-directed mutagenesis at the sites of 244E, 245S, 251S and 252R, with the last 10 amino acids in the amino acid sequence randomly mutated. We evaluated the enzyme activity and substrate specificity. The engineered enzymes showed a relative increase in phosphorylation in the nucleoside analogs of BVDU ((E)-5‑(2-Bromovinyl)-2'-deoxyuridine) or gemcitabine (DFDC, 2',2'-difluoro-deoxycytidine) compared with the wild-type enzyme. The dNKmu enzymes were expressed in the breast cancer cell lines MDA-MB-231 (ER-) and MCF7 (ER+). In studying the sensitivity of the cell lines to DFDC, conditionally replicative adenovirus (CRAd) SG500-dNKmu showed higher expression and enzymatic activity than the replication-defective adenovirus SG500 in cancer cells, but with less cytotoxicity to cancer cells than that of SG500. Our data suggest that the triple phosphorylated DFDC catalyzed by dNKmu inhibited the replication of adenovirus with a simultaneous positive therapeutic effect to cancer cells. Therefore, concomitant use of the SG500‑dNKmu and DFDC could be a novel targeted strategy in suicide gene therapy with safe control of excessive virus replication.

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1.	Zheng X, Johansson M and Karlsson A: Retroviral transduction of cancer cell lines with the gene encoding Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. J Biol Chem. 275:39125–39129. 2000. View Article : Google Scholar : PubMed/NCBI
2.	Zheng X, Johansson M and Karlsson A: Bystander effects of cancer cell lines transduced with the multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster and synergistic enhancement by hydroxyurea. Mol Pharmacol. 60:262–266. 2001.PubMed/NCBI
3.	Zheng X, Johansson M and Karlsson A: Nucleoside analog cytotoxicity and bystander cell killing of cancer cells expressing Drosophila melanogaster deoxyribonucleoside kinase in the nucleus or cytosol. Biochem Biophys Res Commun. 289:229–233. 2001. View Article : Google Scholar : PubMed/NCBI
4.	Betham B, Shalhout S, Marquez VE and Bhagwat AS: Use of Drosophila deoxynucleoside kinase to study mechanism of toxicity and mutagenicity of deoxycytidine analogs in Escherichia coli. DNA Repair (Amst). 9:153–160. 2010. View Article : Google Scholar : PubMed/NCBI
5.	Munch-Petersen B, Knecht W, Lenz C, Sondergaard L and Piskur J: Functional expression of a multisubstrate deoxyribonucleoside kinase from Drosophila melanogaster and its C-terminal deletion mutants. J Biol Chem. 275:6673–6679. 2000. View Article : Google Scholar : PubMed/NCBI
6.	Sokkar P, Sathis V and Ramachandran M: Computational modeling on the recognition of the HRE motif by HIF-1: molecular docking and molecular dynamics studies. J Mol Model. 18:1691–700. 2012. View Article : Google Scholar : PubMed/NCBI
7.	Lee KJ, Lee KY and Lee YM: Downregulation of a tumor suppressor RECK by hypoxia through recruitment of HDAC1 and HIF-1alpha to reverse HRE site in the promoter. Biochim Biophys Acta. 1803:608–616. 2010. View Article : Google Scholar : PubMed/NCBI
8.	Mikkelsen NE, Munch-Petersen B and Eklund H: Structural studies of nucleoside analog and feedback inhibitor binding to Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. FEBS J. 275:2151–2160. 2008. View Article : Google Scholar : PubMed/NCBI
9.	Yu de B, Zhong SY, Yang M, Wang YG, Qian QJ, Zheng S and Liu XY: Potent antitumor activity of double-regulated oncolytic adenovirus-mediated ST13 for colorectal cancer. Cancer Sci. 100:678–683. 2009.PubMed/NCBI
10.	Beltinger C, Fulda S, Kammertoens T, Meyer E, Uckert W and Debatin KM: Herpes simplex virus thymidine kinase/ganciclovir-induced apoptosis involves ligand-independent death receptor aggregation and activation of caspases. Proc Natl Acad Sci USA. 96:8699–8704. 1999. View Article : Google Scholar
11.	Wildner O and Morris JC: The role of the E1B 55 kDa gene product in oncolytic adenoviral vectors expressing herpes simplex virus-tk: assessment of antitumor efficacy and toxicity. Cancer Res. 60:4167–4174. 2000.PubMed/NCBI
12.	Sandrini MP, Clausen AR, On SL, Aarestrup FM, Munch-Petersen B and Piskur J: Nucleoside analogues are activated by bacterial deoxyribonucleoside kinases in a species-specific manner. J Antimicrob Chemother. 60:510–520. 2007. View Article : Google Scholar : PubMed/NCBI
13.	Jordheim LP, Galmarini CM and Dumontet C: Gemcitabine resistance due to deoxycytidine kinase deficiency can be reverted by fruitfly deoxynucleoside kinase, DmdNK, in human uterine sarcoma cells. Cancer Chemother Pharmacol. 58:547–554. 2006. View Article : Google Scholar
14.	Zhang ZL, Zou WG, Luo CX, et al: An armed oncolytic adenovirus system, SG500-gene, demonstrating potent antitumoral efficacy. Cell Res. 13:481–489. 2003. View Article : Google Scholar : PubMed/NCBI
15.	Cappella P, Tomasoni D, Faretta M, et al: Cell cycle effects of gemcitabine. Int J Cancer. 93:401–408. 2001. View Article : Google Scholar : PubMed/NCBI
16.	Morris JC and Wildner O: Therapy of head and neck squamous cell carcinoma with an oncolytic adenovirus expressing HSV-tk. Mol Ther. 1:56–62. 2000. View Article : Google Scholar : PubMed/NCBI
17.	Heilbronn R and Weger S: Viral vectors for gene transfer: current status of gene therapeutics. Handb Exp Pharmacol. 197:143–170. 2010. View Article : Google Scholar : PubMed/NCBI
18.	Nayak S and Herzog RW: Progress and prospects: immune responses to viral vectors. Gene Ther. 17:295–304. 2010. View Article : Google Scholar : PubMed/NCBI
19.	Büning H, Perabo L, Coutelle O, Quadt-Humme S and Hallek M: Recent developments in adeno-associated virus vector technology. J Gene Med. 10:717–733. 2008.

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Conditionally replicating adenovirus SG500-expressed mutant Dm-dNK gene for breast cancer therapy

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