Measurement of γ-H2AX foci, miRNA-101, and gene expression as a means to quantify radiation-absorbed dose in cancer patients who had undergone radiotherapy
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Radiological accidents and nuclear terrorism pose an increased threat to members of the public who, following such an event, would need to be assessed for medical care by fast triage. Assay methods such as chromosome aberrations (CA), cytokinesis-block micronucleus (CBMN) and fluorescence in situ hybridization (FISH) techniques have been well established for dose estimation and their potential for handling more samples has also been proved with automation. However, culturing of lymphocytes is an inevitable step, which limits the potential of these markers for triage. In vitro analysis of gamma-H2AX (γ-H2AX), gene and microRNA (miRNA) markers do not require culturing of lymphocytes, and as such have been suggested as attractive tools for triage. Despite studies reporting in vitro dose–response curves, limited evidence is available evaluating the suitability of these assays in real situations. In this study, we have measured the absorbed dose using γ-H2AX, gene (GADD45A, FDXR, and CDKN1A) and miRNA-101 expression in blood samples of cancer patients (n = 20) who had undergone partial-body radiotherapy and compared with the derived equivalent whole-body doses (EWBD). The obtained results from all patients showed a significant (p < 0.05) increase of γ-H2AX foci in post-irradiated as compared to pre-irradiated samples. Moreover, estimated doses using γ-H2AX foci showed a correlation with the derived EWBD (r2 = 0.60, p = 0.0003) and was also shown to be dependent on the irradiated body volume. Consistent with γ-H2AX foci frequency, an increase in fold change expression of genes and miRNA-101 was observed. However, the estimated dose significantly varied among the subjects and showed poor correlation (r2 = 0.09, 0.04, 0.01 and 0.03 for GADD45A, FDXR, CDKN1A and miRNA-101, respectively) with EWBD. The overall results suggest that the established in vitro γ-H2AX assay is suitable for the detection of radiation exposure and can also provide an estimate of the dose in in vivo irradiated samples. The genes and miRNA-101 markers showed increased expression; nevertheless, there is a need for further improvements to measure doses accurately using these markers.
Keywordsγ-H2AX foci MicroRNA Gene expression Biomarkers Radiotherapy
We acknowledge the support from Medical Physicists Ms. Sidonia Vallas Xavier and nursing staff of Kamakshi Memorial Hospital, Chennai, India, for their help in the collection of blood samples from cancer patients who had undergone radiotherapy. We also acknowledge Dr. Elizabeth A. Ainsbury, Cytogenetics Group Leader, Public Health England Centre for Radiation, Chemical and Environmental Hazards (PHE CRCE), Chilton, Didcot, Oxon OX11 0RQ, UK, for her scientific input.
This work was supported by CSIR (Council of Scientific and Industrial Research), Government of India (File. No. 09/949/0004/2016-EMR-I) and Life Science Research Board, Defense Research Development Organization, Delhi, India, (Ref. No. DLS/81/48222/LSRB-261).
Compliance with ethical standards
Conflict of interest
All the authors declare that there is no conflict of interest.
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