Evaluation of endogenous control gene(s) for gene expression studies in human blood exposed to super(60)Co gamma -rays ex vivo

In gene expression studies, it is critical to normalize data using a stably expressed endogenous control gene in order to obtain accurate and reliable results. However, we currently do not have a universally applied endogenous control gene for normalization of data for gene expression studies, parti...

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Bibliographic Details
Published in:Journal of radiation research 2015-01, Vol.56 (1), p.177-185
Main Authors: Vaiphei, S Thangminlal, Keppen, Joshua, Nongrum, Saibadaiahun, Chaubey, R C, Kma, L, Sharan, R N
Format: Article
Language:English
Subjects:
Algorithms
Assessments
Blood
Females
Gene expression
Genes
Human
Males
Online Access:Get full text
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Summary:In gene expression studies, it is critical to normalize data using a stably expressed endogenous control gene in order to obtain accurate and reliable results. However, we currently do not have a universally applied endogenous control gene for normalization of data for gene expression studies, particularly those involving super(60)Co gamma -ray-exposed human blood samples. In this study, a comparative assessment of the gene expression of six widely used housekeeping endogenous control genes, namely 18S, ACTB, B2M, GAPDH, MT-ATP6 and CDKN1A, was undertaken for a range of super(60)Co gamma -ray doses (0.5, 1.0, 2.0 and 4.0 Gy) at 8.4 Gy min super(-1) at 0 and 24 h post-irradiation time intervals. Using the NormFinder algorithm, real-time PCR data obtained from six individuals (three males and three females) were analyzed with respect to the threshold cycle (Ct) value and abundance, Delta Ct pair-wise comparison, intra- and inter-group variability assessments, etc. GAPDH, either alone or in combination with 18S, was found to be the most suitable endogenous control gene and should be used in gene expression studies, especially those involving qPCR of gamma -ray-exposed human blood samples.
ISSN:0449-3060
1349-9157
DOI:10.1093/jrr/rru074