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Optimization of radiation controlled gene expression by adenoviral vectors in vitro

Cancer Gene Therapy volume 12, pages 640–646 (2005)Cite this article

Abstract

The radiation-inducible EGR-1-promoter has been used in different gene therapy approaches in order to enhance and locally restrict therapeutic efficacy. The aim of this study was to reduce nonspecific gene expression in the absence of irradiation (IR) in an adenoviral vector. Rat rhabdomyosarcoma R1H tumor cells were infected with adenoviral vectors expressing either EGFP or HSV-TK under control of the murine EGR-1 promoter/enhancer. Cells were irradiated at 0–6 Gy. Gene expression was determined by FACS-analysis (EGFP), or crystal violet staining (HSV-TK). The bovine growth hormone polyadenylation signal (BGH pA) was used as insulating sequence and was introduced upstream or upstream and downstream of the expression cassette. Infected R1H cells displayed IR dose-dependent EGFP expression. Cells treated with IR, AdEGR.TK and ganciclovir displayed a survival of 17.3% (6 Gy). However, significant gene expression was observed in the absence of IR with EGR.TK and EGR.EGFP constructs. Introduction of BGHpA upstream or upstream and downstream of expression cassette resulted in decreased nonspecific cytotoxicity by a factor of 1.6–2.3 with minor influence on the induced level of cytotoxicity. Introduction of insulating sequences in adenoviral vectors might allow tighter temporospatial control of gene expression by the radiation-inducible EGR-1 promoter.

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Abbreviations

MOI:

multiplicity of infection

CArG Box:

CC(A/T)6GG like sequence

GCV:

ganciclovir

IR:

irradiation

Egr-1:

early growth response gene

BGH:

bovine growth hormone

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Acknowledgements

The skillful technical assistance of S Wegerer and B Essien is greatly acknowledged. We thank P Wendt for FACS Analysis. Plasmid pE425 was kindly provided by VP Sukhatme. This work was supported by Deutsche Forschungsgemeinschaft (grant number WU 335/1-1). IEOG was supported by a DAAD PhD scholarship.

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Authors and Affiliations

  1. Institut für Experimentelle Onkologie & Therapieforschung, Klinikum rechts der Isar, TU München, Munich, Germany

    Martina Anton, Iman E O Gomaa, Tobias von Lukowicz & Bernd Gansbacher

  2. Klinik und Poliklinik für Strahlentherapie & Radiologische Onkologie, Klinikum rechts der Isar, TU München, Munich, Germany

    Iman E O Gomaa, Tobias von Lukowicz, Michael Molls & Florian Würschmidt

  3. National Center for Radiation Research and Technology, Egyptian Atomic Energy Authority, Cairo, Egypt

    Iman E O Gomaa

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  1. Martina Anton
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Correspondence to Martina Anton.

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Anton, M., Gomaa, I., von Lukowicz, T. et al. Optimization of radiation controlled gene expression by adenoviral vectors in vitro. Cancer Gene Ther 12, 640–646 (2005). https://doi.org/10.1038/sj.cgt.7700829

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