Nitric oxide synthase gene therapy enhances the toxicity of cisplatin in cancer cells
Nitric oxide synthase gene therapy enhances the toxicity of cisplatin in cancer cells
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DOI：
10.1002/jgm.1280
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发表时间：
2009-02-01
影响因子：
3.5
通讯作者：
Hirst, Davi G.
中科院分区：
文献类型：
--
作者：
Adams, Catherine;McCarthy, Helen O.;Hirst, Davi G.
背景一氧化氮（NO中心点）是一种有效的体外化疗增敏剂。我们研究了诱导型一氧化氮合酶（iNOS）基因转移的组合，驱动一个强大的组成型启动子（巨细胞病毒; CMV）与DNA交联剂顺铂在小鼠和人类肿瘤细胞株。方法证明的主要实验中进行的辐射诱导纤维肉瘤-1（RIF-1）小鼠细胞系。在暴露于顺铂之前，使用阳离子脂质载体用组成型表达的CMV/iNOS质粒DNA转染细胞。在皮内RIF-1转移模型中测定体内功效，腹膜内施用顺铂。此外，在各种人肿瘤细胞系中研究了治疗潜力，包括人前列腺癌（DU 145和PC 3）和人结肠癌（HT 29和HCT 116）细胞系。结果CMV/iNOS体外转染RIF-1细胞后，顺铂的细胞毒作用（0.2-1.0 μ M）明显增强。通过直接注射到已建立的RIF-1肿瘤中的CMV/iNOS的体内转移引起了显著（p = 0.0027）的产量生长延迟。CMV/iNOS基因转染后，除HCT 116细胞外，所有细胞均呈强阳性表达，且NO中心点水平均显著升高。结论iNOS高表达导致的NO中心点存在，对顺铂的化疗敏感性显著。我们得出结论，p53状态的各种细胞系是不可能负责顺铂诱导的细胞凋亡。版权所有（C）2008约翰威利父子有限公司
Background Nitric oxide (NO center dot) derived from donor drugs has been shown to be an effective chemosensitizer in vitro. We investigated the combination of inducible nitric oxide synthase (iNOS) gene transfer, driven by a strong constitutive promoter (cytomegalovirus; CMV) with the DNA cross-linking agent cisplatin in mouse and human tumour cell lines.Methods Proof of principal experiments were performed in the radiation-induced fibrosarcoma-1 (RIF-1) murine cell line. Cells were transfected with constitutively expressed CMV/iNOS plasmid DNA using a cationic lipid vector, before exposure to cisplatin. In vivo efficacy was determined in an intradermal RIF-1 turnout model, with intraperitoneal administration of cisplatin. Additionally, treatment potential was investigated in various human turnout cell lines including human prostate (DU145 and PC3) and human colon (HT29 and HCT116) cancer cell lines. Experimental endpoints were established using western blot, Greiss test, clonogenic assay and turnout growth delay.Results Transfection of RIF-1 turnout cells in vitro with the CMV/iNOS significantly enhanced the cytotoxicity of cisplatin (0.2-1.0 PM). In vivo transfer of CMV/iNOS by direct injection into established RIF-1 tumours caused a significant (p = 0.0027) delay in turnout growth. CMV/iNOS gene transfer in vitro resulted in the strong expression of iNOS DNA in all cell lines, and significantly increased levels of NO center dot in all cell lines except HCT116.Conclusions Significant chemosensitization of cisplatin cytotoxicity was observed in the presence of NO center dot derived from the overexpression iNOS. We conclude that p53 status of the various cell lines was unlikely to be responsible for cisplatin-induced apoptosis. Copyright (C) 2008 John Wiley & Sons, Ltd.