- 作者：Petri Nokisalmi ; M.D. ; M.Sc.&lowast ; ; &dagger ; ; Maria Rajecki ; M.D. ; Ph.D.&lowast ; ; &dagger ; ; Sari Pesonen ; Ph.D.&lowast ; ; &dagger ; ; Sophie Escutenaire ; D.V.M. ; Ph.D.&lowast ; ; &dagger ; ; Rabah Soliymani ; M.Sc.&Dagger ; ; Mikko Tenhunen ; Ph.D.§ ; ; Laura Ahtiainen ; Ph.D.&lowast ; ; &dagger ; ; Akseli Hemminki ; M.D. ; Ph.D.&lowast ; ; &dagger ; ; akseli.hemminki@helsinki.fi
- 关键词：Cancer gene therapy ; Adenovirus ; Transgene expression ; Radiotherapy ; DNA repair
- 刊名：International Journal of Radiation Oncology*Biology*Physics
- 出版年：2012
- 出版时间：1 May, 2012
- 年：2012
- 卷：83
- 期：1
- 页码：376-384
- 全文大小：1078 K
Purpose
In the present study, we evaluated the combination of replication-deficient adenoviruses and radiotherapy in?vitro. The purpose of the present study was to analyze the mechanism of radiation-mediated upregulation of adenoviral transgene expression.Methods and Materials
Adenoviral transgene expression (luciferase or green fluorescent protein) was studied with and without radiation in three cell lines: breast cancer M4A4-LM3, prostate cancer PC-3MM2, and lung cancer LNM35/enhanced green fluorescent protein. The effect of the radiation dose, modification of the viral capsid, and five different transgene promoters were studied. The cellular responses were studied using mass spectrometry and immunofluorescence analysis. Double strand break repair was modulated by inhibitors of heat shock protein 90, topoisomerase-I, and DNA protein kinase, and transgene expression was measured.
Results
We found that a wide range of radiation doses increased adenoviral transgene expression regardless of the cell line, transgene, promoter, or viral capsid modification. Treatment with adenovirus, radiation, and double strand break repair inhibitors resulted in persistence of double strand breaks and subsequent increases in adenovirus transgene expression.
Conclusions
Radiation-induced enhancement of adenoviral transgene expression is linked to DNA damage recognition and repair. Radiation induces a global cellular response that results in increased production of RNA and proteins, including adenoviral transgene products. This study provides a mechanistic rationale for combining radiation with adenoviral gene delivery.