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

腺病毒被认为是基因治疗领域中一个很有前途的载体系统，可以将治疗基因传递到靶细胞中。然而，与腺病毒本身有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。观察同时表达2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；其中包括表达针对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 cDNA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erbB2R2/p53中，核酶由pol III启动子驱动，p53同时由CMV启动子驱动。评估了腺病毒载体在几种人类泌尿系统癌细胞系（膀胱癌和前列腺癌）中的感染效果，并标准化了每个细胞系中用于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，在所检查的每个癌细胞中，治疗基因都有充分的表达。采用单独使用和联合使用的方法，对各腺病毒体外抑制细胞生长的效果进行了评价。每种病毒都表现出剂量依赖性的生长抑制。通过将表达核酶的病毒与表达p53的病毒结合，可以观察到协同效应。在双重治疗中，表达基因的病毒，即Ad-HrasRz/p53和Ad-erbB2Rz/p53，在总效价标准的情况下，生长抑制效果优于表达核酶的病毒和表达p53的病毒联合治疗。这些数据表明低滴度双基因表达腺病毒具有足够的治疗效果。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