Medical Radiology and Radiation Safety. 2025. Vol. 70. № 5
DOI:10.33266/1024-6177-2025-70-5-23-27
M.A. Ignatov1, 2, A.K. Chigasova1, 2, 3, A.A. Osipov2, N.Yu. Vorobyeva1, 2,
Yu.A. Fedotov1, 2, Dn.M. Alekseev1, T.I. Gimadova1, А.N. Bashkov1,
Yu.D. Udalov1, A.N. Osipov1, 2
Changes in the Number of Dna Repair Protein Foci in Tomography-Irradiated Human Mesenchymal Stem Cells
1 A.I. Burnazyan Federal Medical Biophysical Center, Moscow, Russia
2 N.N. Semenov Federal Research Center for Chemical Physics, Moscow, Russia
3 Institute of Biochemical Physics, Moscow, Russia
Contact person: A.N. Osipov, e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.
Abstract
Purpose: Comparative analysis of changes in the number of γH2AX, 53BP1 and pATM foci in human mesenchymal stem/stromal cells (MSCs) after irradiation during single and five-time sequential computed tomography (CT). Additionally, as a positive control, changes in these parameters were studied after irradiation of cells using an X-ray machine at a dose of 2000 mGy.
Material and methods: A primary culture of human MSCs was obtained from the collection of BioloT LLC (Russia). A TOSHIBA AQUILION 64 computed tomograph (Japan) was used for cell irradiation. Dosimetric monitoring was carried out using the thermoluminescent method with aluminophosphate dosimeters and magnesium borate-based dosimeters. For comparative studies and obtaining a positive control, a RUST-M1 X-ray biological unit (Russia) equipped with two X-ray emitters (dose 2000 mGy, dose rate 0.85 Gy/min) was used. Immunocytochemical staining using antibodies to γH2AX, 53BP1 and pATM was used for quantitative assessment of γH2AX, 53BP1 and pATM foci. Statistical significance was assessed using analysis of variance (ANOVA).
Results: The studies showed that irradiation of MSCs during CT (absorbed doses during single CT approximetly 88 mGy) causes a statistically significant increase in the number of foci of the proteins γH2AX, 53BP1 and pATM, recorded 0.5 h after irradiation. However, no statistically significant increase in the number of residual γH2AX, 53BP1 and pATM foci was observed 24 h after irradiation compared to the control values. In general, a comprehensive assessment of the number of γH2AX, 53BP1 and pATM protein foci can be recommended for early biodosimetry of irradiation during CT.
Keywords: computed tomography, x-ray radiation, low doses, mesenchymal stem cells, DNA repair, γH2AX, 53BP1, pATM
For citation: Ignatov MA, Chigasova AK, Osipov AA, Vorobyeva NYu, Fedotov YuA, Alekseev DnM, Gimadova TI, Bashkov АN, Udalov YuD, Osipov AN. Changes in the Number of Dna Repair Protein Foci in Tomography-Irradiated Human Mesenchymal Stem Cells. Medical Radiology and Radiation Safety. 2025;70(5):23–27. (In Russian). DOI:10.33266/1024-6177-2025-70-5-23-27
References
1. Han M.A., Kim J.H. Diagnostic X-Ray Exposure and Thyroid Cancer Risk: Systematic Review and Meta-Analysis. Thyroid. 2018;28;2:220-8. doi: 10.1089/thy.2017.0159.
2. Krille L., Dreger S., Schindel R, Albrecht T, Asmussen M, Barkhausen J, et al. Risk of Cancer Incidence before the Age of 15 Years after Exposure to Ionising Radiation from Computed Tomography: Results from a German Cohort Study. Radiat Environ Biophys. 2015;54;1:1-12. doi: 10.1007/s00411-014-0580-3.
3. Kim E.E., Cenzer I., Graham F.J., Kang J., Lee S.J., Rustagi A.S. Time to Benefit for Lung Cancer Screening: a Systematic Review and Survival Meta-Analysis. Am J Prev Med. 2025:107736. doi: 10.1016/j.amepre.2025.107736.
4. Ostrowski M., Marjanski T., Rzyman W. Low-Dose Computed Tomography Screening Reduces Lung Cancer Mortality. Adv Med Sci. 2018;63;2:230-6. doi: 10.1016/j.advms.2017.12.002.
5. Bushmanov A., Vorobyeva N., Molodtsova D., Osipov A.N. Utilization of DNA Double-Strand Breaks for Biodosimetry of Ionizing Radiation Exposure. Environmental Advances. 2022:8. doi: 10.1016/j.envadv.2022.100207.
6. Mah L.J., El-Osta A., Karagiannis T.C. Gamma H2ax: a Sensitive Molecular Marker of DNA Damage and Repair. Leukemia. 2010;24;4:679-86. doi: 10.1038/leu.2010.6.
7. Pustovalova M.V., Nekrasov V.D., Andreev E.V., Fadeikina I.N., Leonov S.V., Nechaev A.N., et al. Synthesized Using β-Cyclodextrin Silver and Gold Nanoparticles as Radiosensitizers in Breast Cancer Radiotherapy. Medical Radiology and Radiation Safety. 2025;70;2:35-9. doi: 10.33266/1024-6177-2025-70-2-35-39.
8. Jakl L., Markova E., Kolarikova L., Belyaev I. Biodosimetry of Low Dose Ionizing Radiation Using DNA Repair Foci in Human Lymphocytes. Genes (Basel). 2020;11:1. doi: 10.3390/genes11010058.
9. Płódowska M., Krakowiak W., Węgierek-Ciuk A., Lankoff A., Szary K., Lis K., et al. Hypothermia Differentially Modulates the Formation and Decay of NBS1, γH2AX and 53BP1 Foci in U2OS Cells Exposed to Gamma Radiation. Scientific Reports. 2022;12:1. doi: 10.1038/s41598-022-09829-y.
10. Osipov A., Chigasova A., Yashkina E., Ignatov M., Vorobyeva N., Zyuzikov N., et al. Early and Late Effects of Low-Dose X-ray Exposure in Human Fibroblasts: DNA Repair Foci, Proliferation, Autophagy, and Senescence. International Journal of Molecular Sciences. 2024;25:15. doi: 10.3390/ijms25158253.
11. Chigasova A.K., Pustovalova M.V., Osipov A.A., Korneva S.A., Eremin P.S., Yashkina E.I., et al. Post-Radiation Changes in The Number of Phosphorylated H2ax and Atm Protein Foci in Low Dose X-Ray Irradiated Human Mesenchymal Stem Cells. Medical Radiology and Radiation Safety. 2024;69;1:15-9. doi: 10.33266/1024-6177-2024-69-1-15-19.
12. Korneva S.A., Chigasova A.K., Osipov A.A., Ignatov M.A., Vorobyova N.Y., Saburov V.O., et al. Post-Irradiation Changes in the Number of γH2ax and Patm Protein Foci in Human Mesenchymal Stem Cells Irradiated with 14.1 MeV Neutrons. Medical Radiology and Radiation Safety. 2025;70;3:11-5. doi: 10.33266/1024-6177-2025-70-3-11-15.
13. Osipov A.A., Chigasova A.K., Yashkina E.I., Ignatov M.A., Vorobyеva N.Y., Osipov A.N. Link between Cellular Senescence and Changes in The Number and Size of Phosphorylated Histone H2ax Foci in Irradiated Human Fibroblasts. Medical Radiology and Radiation Safety. 2024;69;3:13-8. doi: 10.33266/1024-6177-2024-69-3-13-18.
14. Noubissi F.K., McBride A.A., Leppert H.G., Millet L.J., Wang X., Davern S.M. Detection and Quantification of Gamma-H2AX Using a Dissociation Enhanced Lanthanide Fluorescence Immunoassay. Sci Rep. 2021;11;1:8945. doi: 10.1038/s41598-021-88296-3.
15. Prabhu K.S., Kuttikrishnan S., Ahmad N., Habeeba U., Mariyam Z., Suleman M., et al. H2AX: A Key Player in DNA Damage Response and a Promising Target for Cancer Therapy. Biomed Pharmacother. 2024;175:116663. doi: 10.1016/j.biopha.2024.116663.
16. Rass E., Willaume S., Bertrand P. 53BP1: Keeping it Under Control, Even at a Distance from DNA Damage. Genes (Basel). 2022;13:12. doi: 10.3390/genes13122390.
17. Lei T., Du S., Peng Z., Chen L. Multifaceted Regulation and Functions of 53BP1 in NHEJ‑Mediated DSB Repair (Review). Int J Mol Med. 2022;50:1. doi: 10.3892/ijmm.2022.5145.
18. Bartova E., Legartova S., Dundr M., Suchankova J. A Role of the 53BP1 Protein in Genome Protection: Structural and Functional Characteristics of 53BP1-Dependent DNA Repair. Aging (Albany NY). 2019;11;8:2488-511. doi: 10.18632/aging.101917.
19. Shibata A., Jeggo P.A. ATM’s Role in the Repair of DNA Double-Strand Breaks. Genes (Basel). 2021;12:9. doi: 10.3390/genes12091370.
20. Marechal A., Zou L. DNA Damage Sensing by the ATM and ATR Kinases. Cold Spring Harb Perspect Biol. 2013;5:9. doi: 10.1101/cshperspect.a012716.
21. Phan L.M., Rezaeian A.H. ATM: Main Features, Signaling Pathways, and Its Diverse Roles in DNA Damage Response, Tumor Suppression, and Cancer Development. Genes (Basel). 2021;12:6. doi: 10.3390/genes12060845.
22. Osipov A., Chigasova A., Belov O., Yashkina E., Ignatov M., Fedotov Y., et al. Dose Threshold for Residual γH2AX, 53BP1, pATM and p-p53 (Ser-15) Foci in X-ray Irradiated Human Fibroblasts. International Journal of Radiation Biology. 2025;101;3:254-63. doi: 10.1080/09553002.2024.2445581.
23. Gimadova T.I., Keirim-Markus I.B. Experience in Individual Skin Dosimetry at Workplaces and Associated Problems. Radiation Protection Dosimetry. 1991;39;1-3:161-4. doi: 10.1093/oxfordjournals.rpd.a081137.
24. Смирнов В.П., Боженко В.К., Гимадова Т.И., Грабовский Е.В., Грицук А.Н., Иванов А.В. и др. Летальная доза для мышей при облучении импульсным тормозным излучением сверхвысокой мощности дозы на установке Ангара-5-1 // Физика плазмы. 2018. Т.44. №12. С. 1030-5 [Smirnov V.P., Bozhenko V.K., Gimadova T.I., Grabovsky E.V., Gritsuk A.N., Ivanov A.V., et al. Lethal Dose for Mice Irradiated with Pulsed Bremsstrahlung Radiation of Ultra-High Dose Rate at the Angara-5-1 Facility. Fizika Plazmy = Plasma Physics. 2018;44;12:1030-5 (In Russ.)]. doi: 10.1134/s0367292118120132.
25. Osipov A.N., Pustovalova M., Grekhova A., Eremin P., Vorobyova N., Pulin A., et al. Low Doses of X-Rays Induce Prolonged and ATM-independent Persistence of Gamma H2AX foci in Human Gingival Mesenchymal Stem Cells. Oncotarget. 2015;6;29:27275-87. doi: 10.18632/oncotarget.4739.
26. Ingram S.P., Warmenhoven J.W., Henthorn N.T., Chadiwck A.L., Santina E.E., McMahon S.J., et al. A computational Approach to Quantifying Miscounting of Radiation-Induced Double-Strand Break Immunofluorescent Foci. Commun Biol. 2022;5;1:700. doi: 10.1038/s42003-022-03585-5.
27. Pustovalova M., Аstrelina T.A., Grekhova A., Vorobyeva N., Tsvetkova A., Blokhina T., et al. Residual Gamma H2AX foci Induced by Low Dose X-Ray Radiation in Bone Marrow Mesenchymal Stem Cells do not Cause Accelerated Senescence in the Progeny of Irradiated Cells. Aging (Albany NY). 2017;9;11:2397-410. doi: 10.18632/aging.101327.
PDF (RUS) Full-text article (in Russian)
Conflict of interest. The authors declare no conflict of interest.
Financing. The research was supported by the Russian Science Foundation (project No. 23-14-00078).
Contribution. Article was prepared with equal participation of the authors.
Article received: 20.05.2025. Accepted for publication: 25.06.2025.