Lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase driven by the hTERT promoter combined with gemcitabine: A potential strategy for cancer therapy

Zhang,Nianqu; Zhao,Lei; Ma,Shuai; Gu,Ming; Zheng,Xinyu

doi:10.3892/ijmm.2012.1033

Lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase driven by the hTERT promoter combined with gemcitabine: A potential strategy for cancer therapy

Authors:
- Nianqu Zhang
- Lei Zhao
- Shuai Ma
- Ming Gu
- Xinyu Zheng
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Affiliations: Department of Surgical Oncology and Department of General Surgery, The First Affiliated Hospital, China Medical University, Shenyang, P.R. China, Center of Experimental Technology and Medical Research, China Medical University, Shenyang, P.R. China
Published online on: June 14, 2012 https://doi.org/10.3892/ijmm.2012.1033
Pages: 659-665

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Abstract

In contrast to other enzymes, Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) has a broad substrate specificity and high catalytic rate when transferred in human cells. This makes it a promising therapeutic agent when administered together with several cytotoxic nucleoside analogs, such as gemcitabine 2',2'-difluoro-deoxycytidine (dFdC). Therefore, lentiviral vectors, which potentially allow stable long-term transgene expression, are good candidates for gene delivery vehicles. In the present study, we successfully developed a lentivirus-mediated transgene expression system of Dm-dNK under the control of hTERT promoter against the breast cancer cell line (Bcap37), the gastric cancer cell line (SGC7901) and the normal fibroblast cell line (WI-38). Moreover, we also analyzed its targeted cytotoxicity in vitro with treatment of the prodrug dFdC. Bcap37 tumor growth was inhibited in nude mice. Both cancer cell lines exhibited apparent cytotoxicity when infected with recombinant lentivirus constructs expressing Dm-dNK. In contrast, lentivirus-infected WI-38 cells exhibited less cytotoxicity. These data suggested that Dm-dNK was sensitive to dFdC, and it resulted in synergistic growth inhibition and apoptosis induction in vitro. In addition, Lenti-hTERT-dNK/dFdC also suppressed tumor growth in vivo. Our results suggest that the Lenti-hTERT-dNK/dFdC system is a safe and feasible treatment strategy in the development of suicide gene therapy.

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1.	A BertoliM FrancoJ BalzariniM JohanssonA KarlssonAltered deoxyribonucleotide pools in T-lymphoblastoid cells expressing the multisubstrate nucleoside kinase of Drosophila melanogasterFEBS J27239183928200510.1111/j.1742-4658.2005.04808.x16045762
2.	W KnechtNE MikkelsenAR ClausenDrosophila melanogaster deoxyribonucleoside kinase activates gemcitabineBiochem Biophys Res Commun382430433200910.1016/j.bbrc.2009.03.04119285960
3.	S MaL ZhaoZ ZhuThe multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster as a therapeutic suicide gene of breast cancer cellsJ Gene Med133053112011
4.	Z ZhuL MaoL ZhaoSynergistic therapeutic effect in gastric cancer cells produced by oncolytic adenovirus encoding Drosophila melanogaster deoxyribonucleoside kinaseCancer Biol Ther11874882201110.4161/cbt.11.10.1518221383545
5.	M ItoY SudaH HarashimaH KamiyaCytotoxic effect of Drosophila deoxynucleoside kinase gene on replicating plasmid in HeLa cellsBiol Pharm Bull33122312272010
6.	K NishinoT OsakiT KumagaiAdenovirus-mediated gene therapy specific for small cell lung cancer cells using a Myc-Max binding motifInt J Cancer91851856200110.1002/1097-0215(200002)9999:9999%3C::AID-IJC1120%3E3.0.CO;2-111275991
7.	ST YuC LiMH LuNoninvasive and real-time monitoring of the therapeutic response of tumors in vivo with an optimized hTERT promoterCancer11818841893201210.1002/cncr.2647622009660
8.	B YuY ZhangY ZhanCo-expression of herpes simplex virus thymidine kinase and Escherichia coli nitroreductase by an hTERT-driven adenovirus vector in breast cancer cells results in additive antitumor effectsOncol Rep26255264201121573500
9.	S MaW QuL MaoAntitumor effects of oncolytic adenovirus armed with Drosophila melanogaster deoxyribonucleoside kinase in colorectal cancerOncol Rep2714431450201222294034
10.	MP SandriniAR ClausenSL OnFM AarestrupB Munch-PetersenJ PiskurNucleoside analogues are activated by bacterial deoxyribonucleoside kinases in a species-specific mannerJ Antimicrob Chemother60510520200710.1093/jac/dkm24017615154
11.	LM AndersonS SwaminathanI ZackonAK TajuddinB ThimmapayaSA WeitzmanAdenovirus-mediated tissue-targeted expression of the HSVtk gene for the treatment of breast cancerGene Ther6854864199910.1038/sj.gt.330090910505111
12.	Q TangD ZhangM WanL JinExperimental study of the RV-HSV-TK/GCV suicide gene therapy system in gastric cancerCancer Biother Radiopharm22755761200710.1089/cbr.2007.34618158766
13.	JG PanX ZhouR LuoRF HanThe adeno-associated virus-mediated HSV-TK/GCV suicide system: a potential strategy for the treatment of bladder carcinomaMed OncolOct202011(Epub ahead of print)
14.	FQ ZhengY XuRJ YangCombination effect of oncolytic adenovirus therapy and herpes simplex virus thymidine kinase/ganciclovir in hepatic carcinoma animal modelsActa Pharmacol Sin30617627200910.1038/aps.2009.33
15.	M AhnSJ LeeX LiEnhanced combined tumor-specific oncolysis and suicide gene therapy for prostate cancer using M6 promoterCancer Gene Ther167382200910.1038/cgt.2008.5918772902
16.	JF ZhangF WeiHP WangPotent antitumor activity of telomerase-dependent and HSV-TK armed oncolytic adenovirus for non-small cell lung cancer in vitro and in vivoJ Exp Clin Cancer Res2952201010.1186/1756-9966-29-5220487549
17.	X ZhengM JohanssonA KarlssonRetroviral transduction of cancer cell lines with the gene encoding Drosophila melanogaster multisubstrate deoxyribonucleoside kinaseJ Biol Chem2753912539129200010.1074/jbc.M00621220010993893
18.	D TronoLentiviral vectors: turning a deadly foe into a therapeutic agentGene Ther72023200010.1038/sj.gt.330110510680011
19.	M TakakuraS KyoT KanayaCloning of human telomerase catalytic subunit (hTERT) gene promoter and identification of proximal core promoter sequences essential for transcriptional activation in immortalized and cancer cellsCancer Res595515571999
20.	YS CongJ WenS BacchettiThe human telomerase catalytic subunit hTERT: organization of the gene and characterization of the promoterHum Mol Genet8137142199910.1093/hmg/8.1.1379887342
21.	J GuB FangTelomerase promoter-driven cancer gene therapyCancer Biol Ther2Suppl 1S64S70200314508082
22.	W KnechtMP SandriniK JohanssonH EklundB Munch-PetersenJ PiskurA few amino acid substitutions can convert deoxyribonucleoside kinase specificity from pyrimidines to purinesEMBO J2118731880200210.1093/emboj/21.7.187311927571
23.	Z ZhuS MaL ZhaoAdenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase mutants combined with gemcitabine harbor a safe cancer treatment profileInt J Oncol387457532011
24.	C ZhangQT WangH LiuZZ ZhangWL HuangAdvancement and prospects of tumor gene therapyChin J Cancer30182188201110.5732/cjc.010.1007421352695
25.	D Abate-DagaL Garcia-RodriguezL SumoyC FillatCell cycle control pathways act as conditioning factors for TK/GCV sensitivity in pancreatic cancer cellsBiochim Biophys Acta180311751185201010.1016/j.bbamcr.2010.06.00920599444