Abstract：
OBJECTIVES:To develop a novel therapeutic strategy against human bladder cancer using Ad-MK-E1a-a midkine (MK) promoter-regulated, conditionally replicating, adenovirus. METHODS:We tested several human cancer cell lines in vitro, including those of bladder cancer (KK47, 5637, and T24), lung cancer (A549), and head and neck cancer (H891). In each cell line, we examined MK mRNA expression by TaqMan real-time quantitative polymerase chain reaction, MK promoter activity, after plasmid transfection, using a luciferase assay, and the transduction efficiency by co-transfection with the cytomegalovirus-beta-gal plasmid. In these cells, we assessed the cell type-specific replication of Ad-MK-E1a virus by measuring the E1a DNA copy number by real-time polymerase chain reaction and the cell growth inhibition due to this virus using the Alamar blue assay. In animal studies, nude mice were subcutaneously inoculated with KK47 cells and later intratumorally injected with phosphate-buffered saline or Ad5-CMV-LacZ or Ad-MK-E1a. RESULTS:The MK mRNA expression level and MK promoter-driven luciferase activity were relatively greater and markedly increased, respectively, in the 5637, A549, and KK47 cells than in the T24 and H891 cells. After Ad-MK-E1a infection, the E1a DNA copy number increased more significantly in the KK47, 5637, and A549 cells than in the T24 and H891 cells. At a multiplicity of infection of 0.01, Ad-MK-E1a significantly inhibited KK47 and 5637 cell growth. In vivo, Ad-MK-E1a injection markedly inhibited KK47 tumor growth. CONCLUSIONS:We have demonstrated the antitumor effect of Ad-MK-E1a in a human bladder cancer model overexpressing MK mRNA.
journal_name
Urologyjournal_title
Urologyauthors
Terao S,Shirakawa T,Kubo S,Bishunu A,Lee SJ,Goda K,Tsukuda M,Hamada K,Tagawa M,Takenaka A,Fujisawa M,Gotoh Adoi
10.1016/j.urology.2007.07.003subject
Has Abstractpub_date
2007-11-01 00:00:00pages
1009-13issue
5eissn
0090-4295issn
1527-9995pii
S0090-4295(07)01815-8journal_volume
70pub_type
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