| Co-Investigator(Kenkyū-buntansha) |
IWAMURA Masatsugu Kitasato Univ. School of Medicine Assistant Professor, 医学部, 講師 (20176564)
EGAWA Shin Kitasato Univ. School of Medicine Assistant Professor, 医学部, 講師 (60160347)
UCHIDA Toyoaki Kitasato Univ. School of Medicine Assistant Professor, 医学部, 講師 (70146489)
SOH Shigehiro Kitasato Univ. School of Medicine Research Associate, 医学部, 助手 (30206669)
|
|---|
| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| Research Abstract |
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
|
