Drug-resistant human bladder-cancer cells are more sensitive to adenovirus-mediated wild-type p53 gene therapy compared to drug-sensitive cells.
We investigated the therapeutic potential and molecular mechanism of adenovirus-mediated wt p53 gene therapy for drug-resistant human bladder cancers. KK47, a human bladder-cancer cell line, along with the drug-resistant sublines KK47/DDP10, KK47/DDP20 (cisplatin-resistant) and KK47/ADM (doxorubicin-resistant) were used for the experiments. All 4 KK47 cell lines had genetically normal p53 genes. Using an in vitro cytotoxicity assay, the drug-resistant cell lines were more sensitive to Ad-CMV-p53 cell killing than the KK47 parental cell line. Ad-CMV-p53 induced higher levels of p53 protein and mRNA in the drug-resistant cell lines than in the parental cell line and, consequently, higher levels of p21 and Bax mRNA, which resulted in higher percentages of G(1) cell-cycle arrest and apoptosis. The higher efficiencies of adenoviral gene transfer in the drug-resistant cell lines were confirmed by X-gal staining after infection with Ad-CMV-beta-gal. In conclusion, adenovirus-mediated wt p53 gene therapy was more effective in the drug-resistant bladder-cancer cell lines than in the drug-sensitive bladder-cancer cell line.
Shirakawa
T
T
Department of Urology, Kobe University School of Medicine, Kobe, Japan. toshiro@med.kobee-u.ac.jp
Sasaki
R
R
Gardner
T A
TA
Kao
C
C
Zhang
Z J
ZJ
Sugimura
K
K
Matsuo
M
M
Kamidono
S
S
Gotoh
A
A
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D019008:Drug Resistance, Neoplasm; D005091:Exons; D016193:G1 Phase; D016158:Genes, p53; D015316:Genetic Therapy; D006801:Humans; D011518:Proto-Oncogene Proteins / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger / Q000032:analysis; D011991:Receptors, Virus / Q000032:analysis; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000473:pathology / Q000628:therapy*; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D011991:Receptors, Virus; D051028:bcl-2-Associated X Protein