Cytotoxic effects of adenovirus- and lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase on Bcap37 breast cancer cells

Zhang,Nianqu; Dong,Xiaoshen; Sun,Yiqun; Cai,Xiaopeng; Zheng,Caiwei; He,Anning; Xu,Ke; Zheng,Xinyu

doi:10.3892/or.2012.2194

Cytotoxic effects of adenovirus- and lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase on Bcap37 breast cancer cells

Authors:
- Nianqu Zhang
- Xiaoshen Dong
- Yiqun Sun
- Xiaopeng Cai
- Caiwei Zheng
- Anning He
- Ke Xu
- Xinyu Zheng
View Affiliations

Affiliations: Department of Surgical Oncology, Department of General Surgery, First Affiliated Hospital, China Medical University, Shenyang 110001, P.R. China, Department of Burns, General Hospital of Benxi Iron and Steel Company, Benxi, P.R. China, Brandeis University, Waltham, MA, USA, Laboratory 1, Cancer Institute, China Medical University, Shenyang, P.R. China, Department of Radiology, First Affiliated Hospital, China Medical University, Shenyang 110001, P.R. China
Published online on: December 18, 2012 https://doi.org/10.3892/or.2012.2194
Pages: 960-966

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Gene transfer using different viral vectors has demonstrated different antitumor effects in suicide gene therapy. In the present study, in order to optimize the efficacy of replication-defective adenoviral and lentiviral vectors for gene therapy, RT-PCR was used to evaluate the expression of Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) in the Bcap37 human breast cancer cell line, dThd was used to determine the activity of Dm-dNK, cell cytotoxicity was evaluated by MTT assay and cell proliferation was assessed using a hemocytometer. Moreover, apoptosis induction was evaluated by the Annexin V-FITC-labeled FACS method. Furthermore, BALB/C nude mice bearing tumors were treated with Dm-dNK mediated with the pyrimidine nucleoside analog, brivudine [BVDU, (E)-5-(2-bromovinyl)-2'-deoxyuridine]. Our results indicated that the gene expression of Dm-dNK transfected by adenoviral and lentiviral vectors may be detected and that its long-term activity may be retained. Both vectors containing the Dm-dNK gene revealed high cytotoxicity and sensitized cell apoptosis from the BVDU prodrug. In tumor models, lentivirus-mediated gene therapy significantly inhibited the growth of tumors compared with adenovirus-mediated gene therapy. Although adenovirus- and lentivirus-transduced Dm-dNK reveal strong treatment efficacy in vitro, the latter has great potential due to the long-term expression of the therapeutic gene in vivo.

View Figures

View References

1	Akcay MN: Metastatic disease in the breast. Breast. 11:526–528. 2002. View Article : Google Scholar : PubMed/NCBI
2	Munch-Petersen B, Piskur J and Sondergaard L: Four deoxynucleoside kinase activities from Drosophila melanogaster are contained within a single monomeric enzyme, a new multifunctional deoxynucleoside kinase. J Biol Chem. 273:3926–3931. 1998.PubMed/NCBI
3	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
4	Ito M, Suda Y, Harashima H and Kamiya H: Cytotoxic effect of Drosophila deoxynucleoside kinase gene on replicating plasmid in HeLa cells. Biol Pharm Bull. 33:1223–1227. 2010.
5	Zhu Z, Mao L, Zhao L, et al: Synergistic therapeutic effect in gastric cancer cells produced by oncolytic adenovirus encoding Drosophila melanogaster deoxyribonucleoside kinase. Cancer Biol Ther. 11:874–882. 2011. View Article : Google Scholar : PubMed/NCBI
6	Ma S, Zhao L, Zhu Z, et al: The multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster as a therapeutic suicide gene of breast cancer cells. J Gene Med. 13:305–311. 2011.
7	Ma S, Qu W, Mao L, et al: Antitumor effects of oncolytic adenovirus armed with Drosophila melanogaster deoxyribonucleoside kinase in colorectal cancer. Oncol Rep. 27:1443–1450. 2012.PubMed/NCBI
8	Zhang N, Zhao L, Ma S, Gu M and Zheng X: Lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase driven by the hTERT promoter combined with gemcitabine: a potential strategy for cancer therapy. Int J Mol Med. 30:659–665. 2012.
9	Kojaoghlanian T, Flomenberg P and Horwitz MS: The impact of adenovirus infection on the immunocompromised host. Rev Med Virol. 13:155–171. 2003. View Article : Google Scholar : PubMed/NCBI
10	Poller W, Fechner H, Noutsias M, Tschoepe C and Schultheiss HP: Highly variable expression of virus receptors in the human cardiovascular system. Implications for cardiotropic viral infections and gene therapy. Z Kardiol. 91:978–991. 2002. View Article : Google Scholar
11	VandenDriessche T, Collen D and Chuah MK: Viral vector-mediated gene therapy for hemophilia. Curr Gene Ther. 1:301–315. 2001. View Article : Google Scholar : PubMed/NCBI
12	Sugiyama O, An DS, Kung SP, et al: Lentivirus-mediated gene transfer induces long-term transgene expression of BMP-2 in vitro and new bone formation in vivo. Mol Ther. 11:390–398. 2005. View Article : Google Scholar : PubMed/NCBI
13	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
14	Bonini C, Bondanza A, Perna SK, et al: The suicide gene therapy challenge: how to improve a successful gene therapy approach. Mol Ther. 15:1248–1252. 2007. View Article : Google Scholar : PubMed/NCBI
15	Yazawa K, Fisher WE and Brunicardi FC: Current progress in suicide gene therapy for cancer. World J Surg. 26:783–789. 2002. View Article : Google Scholar : PubMed/NCBI
16	Bastide C, Maroc N, Bladou F, et al: Expression of a model gene in prostate cancer cells lentivirally transduced in vitro and in vivo. Prostate Cancer Prostatic Dis. 6:228–234. 2003. View Article : Google Scholar : PubMed/NCBI
17	Johansson M, Brismar S and Karlsson A: Human deoxycytidine kinase is located in the cell nucleus. Proc Natl Acad Sci USA. 94:11941–11945. 1997. View Article : Google Scholar : PubMed/NCBI
18	Zhu C, Johansson M and Karlsson A: Incorporation of nucleoside analogs into nuclear or mitochondrial DNA is determined by the intracellular phosphorylation site. J Biol Chem. 275:26727–26731. 2000.PubMed/NCBI
19	Knecht W, Sandrini MP, Johansson K, Eklund H, Munch-Petersen B and Piskur J: A few amino acid substitutions can convert deoxyribonucleoside kinase specificity from pyrimidines to purines. EMBO J. 21:1873–1880. 2002. View Article : Google Scholar : PubMed/NCBI
20	Solaroli N, Bjerke M, Amiri MH, Johansson M and Karlsson A: Active site mutants of Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. Eur J Biochem. 270:2879–2884. 2003.
21	Zhu Z, Ma S, Zhao L, et al: Adenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase mutants combined with gemcitabine harbor a safe cancer treatment profile. Int J Oncol. 38:745–753. 2011.
22	Knecht W, Mikkelsen NE, Clausen AR, et al: Drosophila melanogaster deoxyribonucleoside kinase activates gemcitabine. Biochem Biophys Res Commun. 382:430–433. 2009. View Article : Google Scholar : PubMed/NCBI
23	Mueller C and Flotte TR: Clinical gene therapy using recombinant adeno-associated virus vectors. Gene Ther. 15:858–863. 2008. View Article : Google Scholar : PubMed/NCBI
24	Calcedo R, Vandenberghe LH, Gao G, Lin J and Wilson JM: Worldwide epidemiology of neutralizing antibodies to adeno-associated viruses. J Infect Dis. 199:381–390. 2009. View Article : Google Scholar : PubMed/NCBI
25	Liu J, Saghizadeh M, Tuli SS, et al: Different tropism of adenoviruses and adeno-associated viruses to corneal cells: implications for corneal gene therapy. Mol Vis. 14:2087–2096. 2008.PubMed/NCBI
26	Sharma A, Tovey JC, Ghosh A and Mohan RR: AAV serotype influences gene transfer in corneal stroma in vivo. Exp Eye Res. 91:440–448. 2010. View Article : Google Scholar : PubMed/NCBI
27	Donsante A, Miller DG, Li Y, et al: AAV vector integration sites in mouse hepatocellular carcinoma. Science. 317:4772007. View Article : Google Scholar : PubMed/NCBI