title: A novel gene expression system for detecting viable bladder cancer cells.
abstract: A novel transcriptional system was developed that can robustly enhance cancer-specific             gene expression. In the system, hTERT promoter-driven gene expression was enhanced             by an advanced two-step transcriptional amplification (TSTA). This construct was             used to develop a novel system for detection of bladder cancer cells. The current             study evaluated the advanced TSTA system by examining the cancer-specific gene             transcription in various bladder cancer cell lines. The system significantly enhanced             cancer-specific luciferase gene expression in the bladder cancer cell lines in             comparison to the previous expression system of one-step or conventional TSTA.             The fold gain of the enhancement was significantly correlated to the telomerase             activity of the cell lines. A green fluorescent protein (GFP) gene encoding plasmid             vector was constructed where hTERT promoter-driving transcription is enhanced             by the advanced TSTA to utilize the system for the imaging and detection of viable             bladder cancer cells. The advanced TSTA-hTERT-GFP plasmid successfully induced             cancer-specific gene expression, showing robust GFP expression in human bladder             cancer cell lines, but no visible GFP expression in normal bladder urothelial             cells. The control GFP plasmid with a CMV promoter yielded GFP expression in both             normal bladder cells and cancer cells. The advanced TSTA-hTERT-GFP plasmid allowed             selective visualization of viable human bladder cancer cells in mixed cell culture             containing 10- and 100-fold more normal bladder urothelial cells. These findings             indicate that the advanced TSTA-hTERT expressional system is a valuable tool for             detecting viable bladder cancer cells. The current system can be applied for in             vitro detection of bladder cancer cells in urine and other types of cancer cells             disseminated in vivo.
authors.affiliation: Center for Gene and Cell Therapy, Okayama University, Okayama                     700-8558, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Ueki
authors.forename: Masami
authors.initials: M
authors.lastname: Watanabe
authors.forename: Haruki
authors.initials: H
authors.lastname: Kaku
authors.forename: Peng
authors.initials: P
authors.lastname: Huang
authors.forename: Shun-Ai
authors.initials: SA
authors.lastname: Li
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Ochiai
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hirata
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Noguchi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamada
authors.forename: Kohji
authors.initials: K
authors.lastname: Takei
authors.forename: Yasutomo
authors.initials: Y
authors.lastname: Nasu
authors.forename: Yuji
authors.initials: Y
authors.lastname: Kashiwakura
authors.forename: Hiromi
authors.initials: H
authors.lastname: Kumon
mesh_terms: D014408:Biomarkers, Tumor / Q000096:biosynthesis / Q000235:genetics*; D045744:Cell Line, Tumor; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases; C509186:TERT protein, human; D019098:Telomerase