Cytotoxic effects of adenovirus- and lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase on Bcap37 breast cancer cells.
Cytotoxic effects of adenovirus- and lentivirus-mediated expression of Drosophila melanogaster deoxyribonucleoside kinase on Bcap37 breast cancer cells.
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DOI：
10.3892/or.2012.2194
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发表时间：
2013-03
期刊：
影响因子：
4.2
通讯作者：
Nianqu Zhang;Xiaoshen Dong;Yiqun Sun;Xiaopeng Cai;Caiwei Zheng;Anning He;Ke-qiang Xu;Xinyu Zheng
中科院分区：
文献类型：
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作者：
Nianqu Zhang;Xiaoshen Dong;Yiqun Sun;Xiaopeng Cai;Caiwei Zheng;Anning He;Ke-qiang Xu;Xinyu Zheng
在自杀基因治疗中，不同病毒载体的基因转移显示出不同的抗肿瘤效果。为了优化复制缺陷腺病毒和慢病毒载体的基因治疗效果，本研究采用RT-PCR法检测黑胃果蝇脱氧核糖核苷激酶（Dm-dNK）在人乳腺癌Bcap37细胞系中的表达，采用dThd法检测Dm-dNK的活性，MTT法检测细胞毒性，血细胞计数仪检测细胞增殖。此外，通过Annexin v - fitc标记的FACS方法评估细胞凋亡诱导。此外，携带肿瘤的BALB/C裸鼠用嘧啶核苷类似物brivudine介导的Dm-dNK治疗[BVDU， (E)-5-(2-bromovinyl)-2'-脱氧尿苷]。我们的结果表明，通过腺病毒和慢病毒载体转染的Dm-dNK可以检测到基因表达，并且可以保持其长期活性。含有Dm-dNK基因的两种载体均显示出高细胞毒性，并使BVDU前药致敏细胞凋亡。在肿瘤模型中，与腺病毒介导的基因治疗相比，慢病毒介导的基因治疗明显抑制肿瘤的生长。虽然腺病毒和慢病毒转导的Dm-dNK在体外显示出较强的治疗效果，但后者由于治疗基因在体内的长期表达而具有很大的潜力。
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.