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Activation of the CMV-IE promoter by hyperthermia in vitro and in vivo: biphasic heat induction of cytosine deaminase suicide gene expression.

Abstract
The cytomegalovirus-immediate early (CMV-IE) promoter is widely used as a strong and constitutively active promoter. Although the CMV-IE promoter does not harbor heat-responsive sequences, we determined its heat inducibility. We analyzed in vitro and in vivo heat responsiveness and possible mechanisms of heat induction of the CMV-IE promoter. We used transfected SW480 human colon carcinoma cells (SW480/CMVCD), expressing CMV-IE promoter-driven bacterial cytosine deaminase (CD) gene. These cells were heated at 42 degrees C. The SW480/CMVCD cells were also used for in vivo studies, in which tumor-bearing animals were treated with hyperthermia at 41.5 degrees C. As controls, SW480 (SW480/HSPCD) cells were used, in which CD expression is driven by the HSP70-promoter. In vitro, we observed a biphasic, up to 25-fold heat induction of CMV-IE-driven CD expression after hyperthermia in SW480/CMVCD cells. In vivo, we found a 2.5-fold induction of CD expression after hyperthermia in SW480/CMVCD tumor-bearing animals. The analysis of the CMV-IE promoter sequence revealed several transcription factor-binding sites, which mediate stress responsiveness. YB-1 and C/EBP-beta might mediate heat responsiveness of the CMV-IE promoter. These data point to limitations in heat-induction gene therapy studies, in which the CMV-IE promoter is used as control system. In addition, the CMV-IE promoter itself could well be used for construction of heat-inducible vectors.
AuthorsDennis Kobelt, Jutta Aumann, Iduna Fichtner, Ulrike Stein, Peter M Schlag, Wolfgang Walther
JournalMolecular biotechnology (Mol Biotechnol) Vol. 46 Issue 2 Pg. 197-205 (Oct 2010) ISSN: 1559-0305 [Electronic] United States
PMID20512535 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Transcription Factors
  • Cytosine Deaminase
Topics
  • Animals
  • Cell Line, Tumor
  • Colonic Neoplasms (genetics, therapy)
  • Cytosine Deaminase (genetics, metabolism)
  • Disease Models, Animal
  • Enzyme Induction (genetics)
  • Gene Expression Regulation, Neoplastic (genetics)
  • Genetic Therapy (methods)
  • Humans
  • Hyperthermia, Induced
  • Male
  • Mice
  • Mice, Inbred NOD
  • Promoter Regions, Genetic (genetics)
  • Transcription Factors (metabolism)

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