Gene therapy using apoptosis-inducer bax and suicide gene for chemically induced rat bladder cancer

使用凋亡诱导剂 bax 和自杀基因治疗化学诱导的大鼠膀胱癌

基本信息

批准号：
12671567
负责人：
SHIBATA Masa-aki
金额：
$ 2.18万
依托单位：
Osaka Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671567/
关键词：
Urinary bladder cancer HSVtk Cancer gene therapy Electroporation Apoptosis Mitochondrial pathway Caspase Non-virus vector Electroporation ミトコンドリア膜電位 Caspase 単純性ヘルペスチミジンキナーゼ bax ラット非ウイルスベクター Electro-gene transfer in vitro /

项目摘要

The effectiveness of in vivo electrogene transfer as a means of gene therapy for rat bladder cancers using the herpes simplex virus 1 thymidine kinase (HSVtk) gene in combination with ganciclovir (GCV) was investigated. In vitro studies demonstrated that 50-70% of the transitional cell carcinoma (TCC) cells died as a result of transfection with pHSVtk and GCV administration and that this treatment was associated with decreased DNA synthesis and elevated activities of caspase-3, -8 and -9. A significantly decreased mitochondrial membrane potential was also noted in TCC cells given pHSVtk+GCV. A direct single injection of HSVtk into bladder tumors using in vivo electrogene transfer followed by GCV administration resulted in significant increases in the levels of apoptosis and histopathological necrosis accompanied by marked inflammation. Active caspase-3 was strongly expressed in the cell death areas of the TCC in rats given pHSVtk/GCV therapy. In vivo electrogene transfer results in efficient gene transfer in chemically induced rat bladder tumors and the HSVtk/GCV producing system induces significant cell death which appears to be, at least, mediated via the mitochondrial apoptotic pathway.

研究了使用单纯疱疹病毒1-胸苷激酶（HSVTK）基因与Ganciclovir（GCV）结合使用的疱疹病毒1-胸苷激酶（HSVTK）基因的大鼠膀胱癌基因治疗的有效性。体外研究表明，过渡细胞癌（TCC）细胞的50-70％因用PHSVTK和GCV给药转染而死亡，并且这种治疗方法与DNA合成降低以及Caspase -3，-8和-9的升高活性有关。给定PHSVTK+GCV，在TCC细胞中还发现了线粒体膜电位的显着降低。直接使用体内电元转移直接将HSVTK注射到膀胱肿瘤中，然后进行GCV给药，从而导致凋亡和组织病理坏死的水平显着增加，并伴有明显的炎症。在PHSVTK/GCV疗法的大鼠中，在TCC的细胞死亡区域中，活性caspase-3强烈表达。体内电orgeene转移导致化学诱导的大鼠膀胱肿瘤和HSVTK/GCV产生系统的有效基因转移，可诱导重要的细胞死亡，这至少是通过线粒体凋亡途径介导的。