Advanced two-step transcriptional amplification as a novel method for cancer-specific gene expression and imaging

Masami Watanabe, Hideo Ueki, Kazuhiko Ochiai, Peng Huang, Yasuyuki Kobayashi, Yasutomo Nasu, Katsumi Sasaki, Haruki Kaku, Yuji Kashiwakura, Hiromi Kumon

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The two-step transcriptional amplification (TSTA) system was previously reported to enhance the tissue-specific gene expression driven by weak promoters, but the enhancement of the gene expression is limited to use in in vitro and in vivo experimental situations. To achieve robust tissuespecific gene expression using the TSTA system, we developed an advanced TSTA system which includes polyglutamines and rat glucocorticoid receptor sequences between the GAL4 and VP16 sequences in the region of the first step of transcription. We evaluated the advanced TSTA system as a method to enhance the human telomerase reverse transcriptase (hTERT) promoter-driving cancer-specific transcription in various cancer cell lines. As a result, the advanced TSTA enhanced cancer-specific luciferase gene expression in all of the examined cancer cell lines, when compared with both the one-step and conventional TSTA systems (an ∼6- and ∼17-fold enhancement, respectively). Notably, the enhancement of the hTERT driven expression by the conventional TSTA system was modest and even inferior to the one-step system in several cancer cell lines. We then constructed a luciferase gene encoding the adenoassociated virus vector in which the hTERT promoter-mediated expression was driven by the advanced TSTA or control systems. In an orthotopic liver tumor model, mice were treated with the vector via tail vein injection. An optical imaging device was used to visualize the in vivo luciferase expression in the orthotopic tumor. The advanced TSTA system significantly enhanced the luciferase expression compared with the one-step and conventional TSTA systems (18.0±1.0- and 15.9±0.85-fold gain, respectively). Therefore, the advanced TSTA system significantly improves hTERT-dependent cancer-specific gene expression both in vitro and in vivo when compared with the previous systems. Since the advanced TSTA method can also be applied to other site-specific gene expression systems using tissue-specific promoters, this approach is expected to become a valuable tool enabling in vivo site-specific targeting in the field of gene therapy and molecular imaging.

Original languageEnglish
Pages (from-to)769-775
Number of pages7
JournalOncology Reports
Volume26
Issue number4
DOIs
Publication statusPublished - Oct 2011

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Neoplasm Genes
Gene Expression
Luciferases
Neoplasms
Cell Line
Optical Devices
Molecular Imaging
Optical Imaging
Glucocorticoid Receptors
Genetic Therapy
Tail
Veins
Viruses
Injections
human TERT protein
Liver
Genes

Keywords

  • Adeno-associated virus
  • Cancer
  • Human telomerase reverse transcriptase
  • Imaging
  • Two-step transcriptional amplification

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Advanced two-step transcriptional amplification as a novel method for cancer-specific gene expression and imaging. / Watanabe, Masami; Ueki, Hideo; Ochiai, Kazuhiko; Huang, Peng; Kobayashi, Yasuyuki; Nasu, Yasutomo; Sasaki, Katsumi; Kaku, Haruki; Kashiwakura, Yuji; Kumon, Hiromi.

In: Oncology Reports, Vol. 26, No. 4, 10.2011, p. 769-775.

Research output: Contribution to journalArticle

Watanabe, Masami ; Ueki, Hideo ; Ochiai, Kazuhiko ; Huang, Peng ; Kobayashi, Yasuyuki ; Nasu, Yasutomo ; Sasaki, Katsumi ; Kaku, Haruki ; Kashiwakura, Yuji ; Kumon, Hiromi. / Advanced two-step transcriptional amplification as a novel method for cancer-specific gene expression and imaging. In: Oncology Reports. 2011 ; Vol. 26, No. 4. pp. 769-775.
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