NDLI: Effect of irradiation on the expression of DNA repair genes studied in human fibroblasts by real-time qPCR using three methods of reference gene validation

Content Provider	Springer Nature Link
Author	Reuther, Sebastian Reiter, Martina Raabe, Annette Dikomey, Ekkehard
Copyright Year	2013
Abstract	The aim of this study was to determine the effects of ionizing radiation on gene expression by using for a first time a qPCR platform specifically established for the detection of 94 DNA repair genes but also to test the robustness of these results by using three analytical methods (global pattern recognition, ΔΔCq/Normfinder and ΔΔCq/Genorm). Study was focused on these genes because DNA repair is known primarily to determine the radiation response. Six strains of normal human fibroblasts were exposed to 2 Gy, and changes in gene expression were analyzed 24 h thereafter. A significant change in gene expression was found for only few genes, but the genes detected were mostly different for the three analytical methods used. For GPR, a significant change was found for four genes, in contrast to the eight or nine genes when applying ΔΔCq/Genorm or ΔΔCq/Normfinder, respectively. When using all three methods, a significant change in expression was only seen for GADD45A and PCNA. These data demonstrate that (1) the genes identified to show an altered expression upon irradiation strongly depend on the analytical method applied, and that (2) overall GADD45A and PCNA appear to play a central role in this response, while no significant change is induced for any of the other DNA repair genes tested.
Starting Page	463
Ending Page	469
Page Count	7
File Format	PDF
ISSN	0301634X
Journal	Radiation and Environmental Biophysics
Volume Number	52
Issue Number	4
e-ISSN	14322099
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2013-07-25
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Irradiation DNA repair Gene expression qPCR MIQE Fibroblasts Biophysics and Biological Physics Effects of Radiation/Radiation Protection Environmental Physics Environmental Monitoring/Analysis Ecosystems
Content Type	Text
Resource Type	Article
Subject	Biophysics Environmental Science Radiation

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