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

腺病毒被认为是一种很有前途的基因治疗载体系统，可以将治疗性基因导入靶细胞。然而，与腺病毒本身相关的不良反应是未来研究的障碍。为了减少腺病毒的数量和提高治疗效果，需要改进载体系统。评价同时表达两种不同治疗基因的腺病毒载体的抗肿瘤效果。设计并重组了几种腺病毒载体，包括表达针对H-ras癌基因的核酶的Ad-HrasRz、表达针对erbB-2基因的核酶的Ad-HrasRz、表达野生型P53的Ad-p53wt、同时表达抗HRAs核酶和P53的Ad-HrasRz/P53以及同时表达抗erbB2核酶和P53的Ad-erbB2Rz/P53。在Ad-HrasRz/P53和ErB2R2/P53中，核酶由PolIII启动子驱动，P53由巨细胞病毒启动子驱动，同时…更狡猾。对腺病毒载体在几种人泌尿系统肿瘤细胞系(膀胱癌和前列腺癌)中的感染效果进行了评价，并对每种细胞系实验的最佳滴度进行了标准化。RT-PCR、Western blotting和免疫细胞化学染色显示治疗基因在每一种癌细胞中都有足够的表达。通过单独使用和联合使用，在体外评价每一种腺病毒抑制细胞生长的效果。每种病毒均表现出剂量依赖性的生长抑制作用。通过核酶表达病毒和p53表达病毒的组合，可以看到协同作用。在双重治疗中，在总滴度标准的情况下，表达病毒的Ad-HrasRz/P53和Ad-erbB2Rz/P53的生长抑制效果优于核酶表达病毒和P53表达病毒的组合。这些数据证明了低滴度双基因表达的腺病毒具有足够的治疗效果。较少

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