PHA stimulation may be useful for FDXR gene expression-based biodosimetry

Document Type : Original Article

Authors

1 Department of Medical Physics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Medical Physics Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Radiology Technology, School of Paramedical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran

4 Immunology Research Center, Inflammation and Inflammatory Diseases Division, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Nowadays, ionizing radiation (IR) has a significant contribution to the diagnostic and therapeutic medicine, and following that, health risks to individuals through unexpected exposure is greatly increased. Therefore, biological and molecular technology for estimation of dose (biodosimetry) is taken into consideration. In biodosimetry methods stimulation of cells to proliferation is routine to achieve more sensitivity of techniques. However, this concept has recently been challenged by new molecular methods such as gene expression analysis. This study aims to investigate the stimulation effects on gene expression biodosimetry.
Materials and Methods: The blood samples were taken from15 patients who were irradiated by TC-99 MIBI, before radiopharmaceutical injection and 24 hr after injection. Lymphocytes were extracted immediately and activated by (phytohemagglutinin) PHA for 24 hr and XPA and FDXR expression levels were investigated by employing relative quantitative Real-Time PCR.
Results: The results of this study show a significant increase in the FDXR expression level and a significant decrease in the XPA after stimulation of irradiated lymphocytes. Interestingly, a significant increasing trend in the FDXR expression level (at 0.05 significance level) following cell stimulation to the division was observed.
Conclusion: Our results suggest that the PHA activation role in gene expression-based biodosimetry is strongly depended on the target genes and the relevant protein pathways. Finally, cell stimulation looks to be useful for some specific genes, such as FDXR, due to the increasing trend in expression and improvement of sensitivity of gene expression-based biodosimetry method.

Keywords


1. Bahreyni-Toossi MT, Najafi-Amiri M, Sankian M, Azimian H, Abdollahi S, Khademi S. INF/IL-4 increases after the low doses of gamma radiation in BALB/c spleen lymphocytes. Iran J Med Phys 2019;16:264-269.
2. Ionizing radiation exposure of the population of the united states. Bethesda, MD: National Council on Radiation Protection and Measurements 1987 NCRP  93.
3. Ionizing radiation exposure of the population of the united states. Bethesda, MD: National Council on Radiation Protection and Measurements 2009 NCRP 160.
4. Khandani AH, Sheikh A. Nuclear medicine.  Clinical Radiation Oncology: Elsevier; 2012. p. 193-201.
5. Eriksson D, Stigbrand T. Radiation-induced cell death mechanisms. Tumor Biol 2010;31:363-372.
6. Fenech M. The lymphocyte cytokinesis-block micronucleus cytome assay and its application in radiation biodosimetry. Health Phys. 2010;98:234-243.
7. Teng Ty, Moffat K. Primary radiation damage of protein crystals by an intense synchrotron X-ray beam. J Synchrotron Radiat 2000;7:313-317.
8. Voisin P. Standards in biological dosimetry: a requirement to perform an appropriate dose assessment. Mutat Res Genet Toxicol Environ Mutagen 2015;793:115-122.
9. Tucker J, Ramsey M, Lee D, Minkler J. Validation of chromosome painting as a biodosimeter in human peripheral lymphocytes following acute exposure to ionizing radiation in vitro. Int J Radiat Biol 1993;64:27-37.
10.    Lin EC, editor Radiation risk from medical imaging. Mayo Clin Proc; 2010: Elsevier.
11.    Guiraud-Vitaux F, Jacquet N, Petiet A, Roy L, Voisin P, Colas-Linhart N. Induction of unstable and stable chromosomal aberrations by 99mTc: in-vitro and in-vivo studies. Nucl Med Commun 2005;26:913-918.
12.    Mogharrabi M, Houman A, Mosaffa N, Tabeie F, Valaei N, Shafiee B. Investigating the effect of high-dose 99mTc-MIBI on chromosomal damage induction among patients undergoing myocardial perfusion scan. Pajoohandeh J 2008;13:9-15.
13.    Tucker JD, Divine GW, Grever WE, Thomas RA, Joiner MC, Smolinski JM, et al. Gene expression-based dosimetry by dose and time in mice following acute radiation exposure. PLoS One 2013;8:e83390.
14.    Santivasi WL, Xia F. Ionizing radiation-induced DNA damage, response, and repair. Antioxid Redox Signal 2014;21:251-259.
15.    Azimian H, Dayyani M, Bahreyni-Toossi MT, Mahmoudi M. Bax/Bcl-2 expression ratio in prediction of response to breast cancer radiotherapy. Iran J Basic Med Sci 2018;21:325-332.
16.    Zhang Y, Rohde LH, Wu H. Involvement of nucleotide excision and mismatch repair mechanisms in double strand break repair. Curr Genomics 2009;10:250-258.
17.    Becker BV, Majewski M, Abend M, Palnek A, Nestler K, Port M, et al. Gene expression changes in human iPSC-derived cardiomyocytes after X-ray irradiation. Int J Radiat Biol 2018;94:1095-1103.
18.    Knops K, Boldt S, Wolkenhauer O, Kriehuber R. Gene expression in low-and high-dose-irradiated human peripheral blood lymphocytes: possible applications for biodosimetry. Radiat Res 2012;178:304-312.
19.    O’Brien G, Cruz-Garcia L, Majewski M, Grepl J, Abend M, Port M, et al. FDXR is a biomarker of radiation exposure in vivo. Sci Rep 2018;8:684.
20.    Fachin AL, Mello SS, Sandrin-Garcia P, Junta CM, Ghilardi-Netto T, Donadi EA, et al. Gene expression profiles in radiation workers occupationally exposed to ionizing radiation. J Radiat Res 2009;50:61-71.
21.    Paul S, Amundson SA. Development of gene expression signatures for practical radiation biodosimetry. Int J Radiat Oncol Biol Phys 2008;71:1236-1244.
22.    Boldt S, Knops K, Kriehuber R, Wolkenhauer O. A frequency-based gene selection method to identify robust biomarkers for radiation dose prediction. Int J Radiat Biol 2012;88:267-276.
23.    Beinke C, Port M, Ullmann R, Gilbertz K, Majewski M, Abend M. Analysis of gene expression changes in PHA-M stimulated lymphocytes–unraveling PHA activity as prerequisite for dicentric chromosome analysis. Rad Res 2018;189:579-596.
24.    Pearton S, Ren F, Patrick E, Law M, Polyakov AY. Ionizing radiation damage effects on GaN devices. ECS J Solid State Sci Technol 2016;5:35-60.
25.    Einor D, Bonisoli-Alquati A, Costantini D, Mousseau T, Møller A. Ionizing radiation, antioxidant response and oxidative damage: a meta-analysis. Sci Total Environ 2016;548:463-471.
26.    Azzam EI, Jay-Gerin JP, Pain D. Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury. Cancer Lett 2012;327:48-60.
27.    Tang FR, Loke WK. Molecular mechanisms of low dose ionizing radiation-induced hormesis, adaptive responses, radioresistance, bystander effects, and genomic instability. Int J Radiat Biol 2015;91:13-27.
28.    Azimian H, Bahreyni-Toossi MT, Rezaei AR, Rafatpanah H, Hamzehloei T, Fardid R. Up-regulation of Bcl-2 expression in cultured human lymphocytes after exposure to low doses of gamma radiation. J Med Phys 2015;40:38-44.
29.    Bahreyni-Toossi M, Azimian H, Rezaei A, Rafatpanah H, Hamzehloei T, Fardid R, editors. Low-dose irradiation alters the radio-sensitivity of human peripheral blood lymphocytes. World Congress on Medical Physics and Biomedical Engineering May 26-31, 2012, Beijing, China; 2013: Springer.
30.    Fliedner TM, Graessle DH, Meineke V, Feinendegen LE. Hemopoietic response to low dose-rates of ionizing radiation shows stem cell tolerance and adaptation. Dose Response 2012;10:644–663.
31.    Hendrick RE. Radiation doses and cancer risks from breast imaging studies. Radiol 2010;257:246-253.
32.    Roy L, Gruel G, Vaurijoux A. Cell response to ionising radiation analysed by gene expression patterns. Ann Ist Super Sanita 2009;45:272-277.
33.    Zhang Y, Rohde LH, Emami K, Hammond D, Casey R, Mehta SK, et al. Suppressed expression of non-DSB repair genes inhibits gamma-radiation-induced cytogenetic repair and cell cycle arrest. DNA Repair 2008;7:1835-1845.
34.    Budworth H, Snijders AM, Marchetti F, Mannion B, Bhatnagar S, Kwoh E, et al. DNA repair and cell cycle biomarkers of radiation exposure and inflammation stress in human blood. PLoS One 2012;7:e48619.
35.    Bahreyni-Toossi MT, Vosoughi H, Azimian H, Rezaei AR, Momennezhad M. In vivo exposure effects of 99mTc-methoxyisobutylisonitrile on the FDXR and XPA genes expression in human peripheral blood lymphocytes. Asia Ocean J Nucl Med Biol 2018;6:32-40.
36.    Mayer C, Popanda O, Zelezny O, von Brevern M-C, Bach A, Bartsch H, et al. DNA repair capacity after γ-irradiation and expression profiles of DNA repair genes in resting and proliferating human peripheral blood lymphocytes. DNA Repair 2002;1:237-250.
37.    Karachristou I, Karakosta M, Pantelias A, Hatzi VI, Karaiskos P, Dimitriou P, et al. Triage biodosimetry using centromeric/telomeric PNA probes and Giemsa staining to score dicentrics or excess fragments in non-stimulated lymphocyte prematurely condensed chromosomes. Mutat Res Genet Toxicol Environ Mutagen 2015;793:107-114.
38.    Soni A, Murmann-Konda T, Magin S, Iliakis G. A method for the cell-cycle-specific analysis of radiation-induced chromosome aberrations and breaks. Mutat Res Fundam Mol Mech Mutagen 2019;815:10-19.
39.    Abend M, Badie C, Quintens R, Kriehuber R, Manning G, Macaeva E, et al. Examining radiation-induced in vivo and in vitro gene expression changes of the peripheral blood in different laboratories for biodosimetry purposes: first RENEB gene expression study. Radiat Res 2016;185:109-123.
40.    Paul S, Amundson SA. Development of gene expression signatures for practical radiation biodosimetry. Int J Radiat Oncol Biol Phys 2008;71:1236-1244. e76.
41.    Rohatgi A. WebPlotDigitizer: Web based tool to extract data from plots, images, and maps. Retrived from http://arohatgi info/WebPlotDigitizer/citation html 2016.
42.    Beinke C, Port M, Ullmann R, Gilbertz K, Majewski M, Abend M. Analysis of gene expression changes in PHA-M stimulated lymphocytes–unraveling PHA activity as prerequisite for dicentric chromosome analysis. Radiat Res 2018;189:579-596.
43.    Hersh EM, Patt YZ, Murphy SG, Dicke K, Zander A, Adegbite M, et al. Radiosensitive, thymic hormone-sensitive peripheral blood suppressor cell activity in cancer patients. Cancer Res 1980;40:3134-3140.
44.    Bryant PE, Riches AC, Shovman O, Dewar JA, Adamson DJ. Topoisomerase IIα levels and G2 radiosensitivity in T-lymphocytes of women presenting with breast cancer. Mutagenesis 2012;27:737-741.