Gene Expression Biodosimetry: Quantitative Assessment of Radiation Dose with Total Body Exposure of Rats

Abstract

Background: Accurate dose assessment and correct identification of irradiated from non-irradiated people are goals of biological dosimetry in radiation accidents. Objectives: Changes in the FDXR and the RAD51 gene expression (GE) levels were here analyzed in response to total body exposure (TBE) to a 6 MV x-ray beam in rats. We determined the accuracy for absolute quantification of GE to predict the dose at 24 hours. Materials and Methods: For this in vivo experimental study, using simple randomized sampling, peripheral blood samples were collected from a total of 20 Wistar rats at 24 hours following exposure of total body to 6 MV X-ray beam energy with doses (0.2, 0.5, 2 and 4 Gy) for TBE in Linac Varian 2100C/D (Varian, USA) in Golestan Hospital, in Ahvaz, Iran. Also, 9 rats was irradiated with a 6MV X-ray beam at doses of 1, 2, 3 Gy in 6MV energy as a validation group. A sham group was also included. After RNA extraction and DNA synthesis, GE changes were measured by the QRT-PCR technique and an absolute quantification strategy by taqman methodology in peripheral blood from rats. ROC analysis was used to distinguish irradiated from non-irradiated samples (qualitative dose assessment) at a dose of 2 Gy. Results: The best fits for mean of responses were polynomial equations with a R2 of 0.98 and 0.90 (for FDXR and RAD51 dose response curves, respectively). Dose response of the FDXR gene produced a better mean dose estimation of irradiated "validation" samples compared to the RAD51 gene at doses of 1, 2 and 3 Gy. FDXR gene expression separated the irradiated rats from controls with a sensitivity, specificity and accuracy of 87.5%, 83.5% and 81.3%, respectively, 24 hours after dose of 2 Gy. These values were significantly (p<0.05) higher than the 75%, 75% and 75%, respectively, obtained using gene expression of RAD51 analysis at a dose of 2 Gy. Conclusions: Collectively, these data suggest that absolute quantification by gel purified quantitative RT-PCR can be used to measure the mRNA copies for GE biodosimetry studies at comparable accuracy to similar methods. In the case of TBE with 6MV energy, FDXR gene expression analysis is more precise than that with RAD51 for quantitative and qualitative dose assessment.

Serum Proteomic and Oxidative Modification Profiling in Mice Exposed to Total Body X-Irradiation

“…For MRM-HR profiling of mOMSA, peptides containing each oxidatively modified amino acid residue were standardized against the peak area value of the corresponding unmodified peptide and analyzed for correlation with the radiation exposure dose as a relative peak area ratio. The fitting of quadratic equations was investigated for each mOMSA; in fact, most human genes show quadratic dose response to radiation [ 16 , 17 ]. In the in vivo model, eleven sequences showed significant dose-dependent correlations ( r value) of >0.5 by linear or curve fitting.…”

Early molecular markers for retrospective biodosimetry and prediction of acute health effects

“…(continued.) methodology), teams used only 1-4 genes [54,[64][65][66][67][68][69][70][71][72][73]. Large scale inter-laboratory comparison exercises initiated from NATO research task groups or the European biodosimetry network (RENEB) confirmed the applicability of this approach [74][75][76][77].…”

“…IFN-gamma) replaced these so called 'immediate early (proto-onco) genes' [50][51][52][53]. GADD45 (Growth arrest and DNA damage gene [54]) was one of the earliest TP53 related genes and many more were examined in the following decades [50].…”

“…from leukaemia, prostate, breast cancer or receiving a computer tomography (CT-scan). Two animal studies in baboons and rats are reported [54,66]. A methodological validation on the same samples using microarrays and qRT-PCR combined was performed in the majority of studies, but an independent validation using different samples was reported for six studies.…”

“…Many laboratories worldwide established their own gene set comprising 1-4 genes for dose estimation using qRT-PCR [54,[64][65][66][67][68][69][70][71][72][73]. Dose estimation requires a calibration curve comprising irradiated samples examined at the same time after irradiation as the potentially irradiated samples of some radiation victims.…”