A novel gene expression system for detecting viable bladder cancer cells

Hideo Ueki, Masami Watanabe, Haruki Kaku, Peng Huang, Shun Ai Li, Kazuhiko Ochiai, Takeshi Hirata, Hirofumi Noguchi, Hiroshi Yamada, Kohji Takei, Yasutomo Nasu, Yuji Kashiwakura, Hiromi Kumon

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)135-140
Number of pages6
JournalInternational Journal of Oncology
Volume41
Issue number1
DOIs
Publication statusPublished - Jul 2012

Fingerprint

Urinary Bladder Neoplasms
Green Fluorescent Proteins
Gene Expression
Neoplasm Genes
Plasmids
Cell Line
Urinary Bladder
Telomerase
Luciferases
Neoplasms
Cell Culture Techniques
Urine

Keywords

  • Bladder cancer
  • Green fluorescent protein
  • Human telomerase reverse transcriptase
  • Plasmid
  • Two-step transcriptional amplification

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

A novel gene expression system for detecting viable bladder cancer cells. / Ueki, Hideo; Watanabe, Masami; Kaku, Haruki; Huang, Peng; Li, Shun Ai; Ochiai, Kazuhiko; Hirata, Takeshi; Noguchi, Hirofumi; Yamada, Hiroshi; Takei, Kohji; Nasu, Yasutomo; Kashiwakura, Yuji; Kumon, Hiromi.

In: International Journal of Oncology, Vol. 41, No. 1, 07.2012, p. 135-140.

Research output: Contribution to journalArticle

Ueki, Hideo ; Watanabe, Masami ; Kaku, Haruki ; Huang, Peng ; Li, Shun Ai ; Ochiai, Kazuhiko ; Hirata, Takeshi ; Noguchi, Hirofumi ; Yamada, Hiroshi ; Takei, Kohji ; Nasu, Yasutomo ; Kashiwakura, Yuji ; Kumon, Hiromi. / A novel gene expression system for detecting viable bladder cancer cells. In: International Journal of Oncology. 2012 ; Vol. 41, No. 1. pp. 135-140.
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