权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Anti-tumor efficacy of the adenoviral vectors those express anti-oncogene ribozyme and wild-type p53 simultaneously

同时表达抗癌基因核酶和野生型p53的腺病毒载体的抗肿瘤功效

基本信息

批准号：
11671580
负责人：
IRIE Akira
金额：
$ 1.6万
依托单位：
Kitasato University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

Adenovirus is thought to be a promising vector system in the field of gene therapy for delivering the therapeutic genes into the targeted cells. However, the adverse effects related to adenoviruses themselves are the obstacles for future studies. Improvement of the vector system is desired for reducing the amount of adenoviruses and for enhancing the therapeutic efficacy. The anti-tumor efficacy of adenoviral vectors those express 2 different therapeutic genes simultaneously, were evaluated. Several kinds of adenoviral vector were designed and recombined; those include Ad-HrasRz which expresses ribozyme against the H-ras oncogene, Ad-HrasRz which expresses ribozyme against the erbB-2 gene, Ad-p53wt which expresses the wild-type p53 cDNA, Ad-HrasRz/p53 which expresses both anti-Hras ribozyme and p53, and Ad-erbB2Rz/p53 which expresses both anti-erbB2 ribozyme and p53. In Ad-HrasRz/p53 and erbB2R2/p53, ribozyme was driven by pol III promoter and p53 was driven by CMV promoter simultaneou … More sly. Infection efficacy of the adenoviral veotor in several human urologic cancer cell lines (bladder cancer and prostate cancer) was evaluated, and the optimal titer of adenovirus for experiments in each cell line was standardized. RT-PCR, western blotting, and immunocytochemical staining demonstrated sufficient expression of the therapeutic genes in every cancer cells examine. The efficacy of cell growth suppression of each adenovirus was evaluated in vitro by solitary uses and by combination. Each virus demonstrated the growth suppression in a dose-dependent manner. The synergistic effect was seen by combination of a ribozyme-expressing virus and a p53-expressing virus. In the dual therapeutic, genes, expressing viruses, I.e., Ad-HrasRz/p53 and Ad-erbB2Rz/p53, growth, suppression efficacy was superior to the combination of ribozyme-expressing virus and p53-expressing virus, when the total amount of titer was standard. These data demonstrated the sufficient therapeutic efficacy of dual genes-expressing adenoviruses with lower-titers. Less

腺病毒是基因治疗领域中一种很有前途的载体系统，可以将治疗基因导入靶细胞。然而，腺病毒本身的副作用是未来研究的障碍。为了减少腺病毒的量和提高治疗效果，需要改进载体系统。同时表达两种不同治疗基因的腺病毒载体的抗肿瘤效果进行了评估。设计并重组了几种腺病毒载体，包括表达抗H-ras癌基因核酶的Ad-HrasRz、表达抗erbB-2基因核酶的Ad-HrasRz、表达野生型p53 cDNA的Ad-p53 wt、同时表达抗Hras核酶和p53的Ad-HrasRz/p53、同时表达抗erbB-2核酶和p53的Ad-erbB 2 Rz/p53。在Ad-HrasRz/p53和erbB 2 R2/p53中，pol III启动子驱动核酶，CMV启动子驱动p53， ...更多信息狡猾评价腺病毒载体在几种人泌尿系统癌细胞系（膀胱癌和前列腺癌）中的感染效力，并标准化用于在每种细胞系中进行实验的腺病毒的最佳滴度。RT-PCR、Western blotting和免疫细胞化学染色显示，治疗基因在每一个检测的癌细胞中都有足够的表达。通过单独使用和组合使用，在体外评价每种腺病毒的细胞生长抑制功效。每种病毒均以剂量依赖性方式表现出生长抑制。通过核酶表达病毒和p53表达病毒的组合观察到协同效应。在双重治疗中，表达病毒的基因，即，在总滴度为标准的情况下，Ad-HrasRz/p53和Ad-erbB 2 Rz/p53的生长抑制效果上级核酶表达病毒和p53表达病毒的联合。这些数据证明了具有较低滴度的双基因表达腺病毒的足够的治疗功效。少