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Cancer Research Institute, Tomsk National Research Medical Center, Russian Academy of Sciences (Junior Researcher)
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Russian Federation
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Siberian State Medical University (Associate Professor)
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Cancer Research Institute, Tomsk National Research Medical Center, Russian Academy of Sciences
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National Research Tomsk State University
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Siberian State Medical University (Professor)
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Purpose: To study the relationship between the methylation status of the Bаk1 promoter and the frequency of chromosomal aberrations in human blood lymphocytes under chronic low-intensity external ionizing radiation. Material and methods: The study was performed on 41 people (31 men and 10 women, aged from 36 to 83 years) who are former or current employees of the Siberian Group of Chemical Enterprises, who have been exposed or haven't been exposed to chronic low-intensity external radiation in the course of their professional activities. The workers included in the study were divided into two groups: the first – 15 people who did not have exposure, the second – 26 people who had external exposure (gamma radiation, total dose 89–716 mSv). Whole blood was used to isolate DNA and evaluate chromosomal aberrations in lymphocytes. The methylation status of the Bаk1 promoter was determined using methylsensitive PCR, which was performed after pretreatment of the isolated DNA with methylsensitive AoxI restrictase. The obtained quantitative data were analyzed using the Statistica 10. The differences were considered statistically significant at p ≤ 0.05. Results: The methylation status of the Bаk1 promoter in the study groups does not differ (p = 0.18). The study of the effect of external radiation dose on the methylation status of the Bak1 promoter showed that the unmethylated promoter prevails in workers with an average radiation dose of 273.37 ± 43.82 mSv, while the methylated promoter – in workers with an average radiation dose of 183.63 ± 20.58 mSv (p = 0.03). The unmethylated promoter Bаk1 is associated with an increased frequency of chromatid fragments in the blood lymphocytes of group 2 workers (p = 0.03). Conclusion: The status of methylation of the Bаk1 promoter in human blood lymphocytes under chronic low-intensity ionizing radiation does not change, but it is observed to depend on the radiation dose and is associated with an increased frequency of chromosomal aberrations (chromatid fragments). Thus, the unmethylated Bаk1 promoter prevails when the external radiation dose is increased. An increase in the frequency of chromatid fragments is associated with an unmethylated Bаk1 promoter. In addition, exposure to chronic low-intensity external radiation is accompanied by a decrease in the frequency of chromosomal fragments in the blood lymphocytes of workers of the Siberian Group of Chemical Enterprises.
chromosomal aberrations, methylation of promoters, apoptosis, chronic low-intensity external irradiation
1. Mazurik VK Rol' regulyatornyh setey otveta kletok na povrezhdeniya v formirovanii radiacionnyh effektov. Radiacionnaya biologiya. Radioekologiya. 2005;45(1):26-45. [Mazurik VK The role of regulatory networks of cell response to damage in the formation of radiation effects. Radiation Biology. Radioecology. 2005;45(1):26-45. (In Russ.)].
2. Varga OYu, Ryabkov VA Apoptoz: ponyatie, mehanizmy realizacii, znachenie. Ekologiya cheloveka. 2006;(7):28-32. [Varga OYu, Ryabkov VA. Apoptosis: concept, mechanisms of implementation, meaning. Human Ecology. 2006;(7):28-32. (In Russ.)].
3. Grabovskiy EV, Oleynik GM, Krastelev EG, Smirnov VP, Hmelevskiy EV, Bozhenko VK i dr. Analiz indukcii apoptoza limfocitov perifericheskoy krovi cheloveka sverhintensivnym gamma-izlucheniem in vitro. Vestnik Rossiyskogo gosudarstvennogo medicinskogo universiteta. 2017;(6):59-66. [Grabovsky EV, Oleinik GM, Krastelev EG, Smirnov VP, Khmelevsky EV, Bozhenko VK, et al. Analysis of induction of apoptosis of human peripheral blood lymphocytes by super-intense gamma radiation in vitro. Bulletin of the Russian State Medical University. 2017;(6):59-66. (In Russ.)].
4. Litvyakov NV, Karpov AB, Tahauov RM, Freydin MB, Sazonov AE, Halyuzova MV Geneticheskie markery individual'noy radiochuvstvitel'nosti cheloveka. Tomsk: Izd-vo Tom. un-ta. 2011. 186 s. [Litvyakov NV, Karpov AB, Takhauov RM, Freydin MB, Sazonov AE, Khalyuzova MV Genetic markers of individual human radio sensitivity. Tomsk: Tomsk University Press; 2011. p. 186. (In Russ.)].
5. Oakes SA, Scorrano L, Opferman JT, Bassik MC, Nishino M, Pozzan T, et al. Proapoptotic BAX and BAK regulate the type 1 inositol trisphosphate receptor and calcium leak from the endoplasmic reticulum. Proceedings of the National Academy of Sciences. 2005;102(1):105-10. DOI:https://doi.org/10.1073/pnas.0408352102.
6. Struijk L, van der Meijden E, Kazem S, ter Schegget J, de Gruijl FR, Steenbergen RDM, et al. Specific betapapillomaviruses associated with squamous cell carcinoma of the skin inhibit UVB-induced apoptosis of primary human keratinocytes. Journal of General Virology. 2008;89(9):2303-14. DOI:https://doi.org/10.1099/vir.0.83317-0.
7. Cartron P-F, Juin P, Oliver L, Martin S, Meflah K, Vallette FM Nonredundant role of Bax and Bak in bid-mediated apoptosis. Molecular and Cellular Biology. 2003;23(13):4701-12. DOI:https://doi.org/10.1128/MCB.23.13.4701-4712.2003.
8. Choi SY, Kim MJ, Kang CM, Bae S, Cho CK, Soh JW, et al. Activation of Bak and Bax through c-Abl-protein kinase Cδ-p38 MAPK signaling in response to ionizing radiation in human non-small cell lung cancer cells. Journal of Biological Chemistry. 2006;281(11):7049-59. DOI:https://doi.org/10.1074/jbc.M512000200.
9. Pendina AA, Grinkevich VV, Kuznecova TV, Baranov VS Metilirovanie DNK - universal'nyy mehanizm regulyacii aktivnosti genov. Ekologicheskaya genetika. 2004;2:27-8. [Pendina AA, Grinkevich VV, Kuznetsova TV, Baranov VS. DNA Methylation - a universal mechanism for regulating gene activity. Ecological Genetics. 2004;2:27-8. (In Russ.)].
10. Mathios D, Hwang T, Xia Y, Phallen J, Rui Y, See AP, et al. Genome-wide investigation of intragenic DNA methylation identifies ZMIZ1 gene as a prognostic marker in glioblastoma and multiple cancer types. International Journal of Cancer. 2019;145(12):3425-35. DOI:https://doi.org/10.1002/ijc.32587.
11. Qiu Y, Gao Y, Yu D, Zhong L, Cai W, Ji J, et al. Genome-wide analysis reveals zinc transporter ZIP9 regulated by DNA methylation promotes radiation-induced skin fibrosis via the TGF-β signaling pathway. Journal of Investigative Dermatology. 2020;140(1):94-102.e7. DOI:https://doi.org/10.1016/j.jid.2019.04.027.
12. Petrenko A, Eshilev E, Kisseljova N Identification of CpG islands hypermethylated in tumor and transformed cells by the methylation-sensitive arbitrarily primed polymerase chain reaction. Molecular Biology. 2012;34(3):408-12. DOI:https://doi.org/10.1007/BF02759673.
13. Kuz'mina NS, Myazin AE, Lapteva NSh, Rubanovich AV Izuchenie aberrantnogo metilirovaniya v leykocitah krovi likvidatorov avarii na ChAES. Radiacionnaya biologiya. Radioekologiya. 2014;54(2):127-39. [Kuzmina NS, Myazin AE, Lapteva NS, Rubanovich AV Study of aberrant methylation in blood leukocytes of liquidators of the chernobyl accident. Radiation Biology. Radioecology. 2014;54(2):127-39. (In Russ.)].
14. 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. Journal of Medical Physics. 2015;40(1):38-44. DOI:https://doi.org/10.4103/0971-6203.152249.
15. Mirabello L, Kratz CP, Savage SA, Greene MH Promoter methylation of candidate genes associated with familial testicular cancer. International Journal of Molecular Epidemiology and Genetics. 2012;3(3):213-27.