mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2023.051mes-42Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕORIGINAL RESEARCHМетилирование промоторов генов клеточного цикла и апоптоза у облученных лиц, впоследствии заболевших злокачественными новообразованиямиMethylation of cell cycle and apoptosis genes’ promoters in exposed individuals with subsequent malignant neoplasmsБлиноваЕ. А.BlinovaE. A.

Евгения Андреевна Блинова

ул. Воровского, д. 68А, г. Челябинск, 154141

Chelyabinsk

КореченковаА. В.KorechenkovaA. V.

Челябинск

Chelyabinsk

НикифоровВ. С.NikiforovV. S.

Челябинск

Chelyabinsk

АклеевА. В.AkleyevA. V.

Челябинск

Chelyabinsk

Уральский научно-практический центр радиационной медицины Федерального медико-биологического агентства России; Челябинский государственный университетРоссияUrals Research Center for Radiation Medicine; Chelyabinsk State UniversityRussian FederationУральский научно-практический центр радиационной медицины Федерального медико-биологического агентства РоссииРоссияUrals Research Center for Radiation MedicineRussian FederationУральский научно-практический центр радиационной медицины Федерального медико-биологического агентства России; Челябинский государственный университетРоссияUrals Research Center for Radiation MedicineRussian Federation2023221020242546369Copyright © Блинова Е.А., Кореченкова А.В., Никифоров В.С., Аклеев А.В., 20242024Блинова Е.А., Кореченкова А.В., Никифоров В.С., Аклеев А.В.Blinova E.A., Korechenkova A.V., Nikiforov V.S., Akleyev A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.extrememedicine.ru/jour/article/view/42

Метилирование ДНК играет важную роль в канцерогенезе, в литературе встречается достаточно много исследований уровня метилирования всего генома, промоторов генов и некодирующих элементов в раковых клетках. При этом данных об изменении паттерна метилирования в клетках крови и связи с развитием злокачественных новообразований (ЗНО) существенно меньше. Цель работы — исследование уровня метилирования промоторных регионов генов контроля клеточного цикла и апоптоза (BAX, MDM2, TP53, NFkB1) в клетках периферической крови лиц, подвергшихся хроническому радиационному воздействию в латентном периоде развития злокачественных новообразований. Исследование проводили у 200 человек, подвергшихся аварийному хроническому радиационному воздействию в результате сбросов радиоактивных отходов в реку Течу. Уровень метилирования оценивали методом ПЦР в реальном времени. Было установлено, что распределение облученных лица с ЗНО в латентном периоде по уровню метилирования промоторных регионов генов BAX, MDM2 и NFkB1 статистически значимо отличалось от распределения в группы сравнения (р < 0,001; р < 0,001; р = 0,004 соответственно). Установлено, что в группе облученных лиц, которые впоследствии заболели ЗНО, доля лиц с уровнем метилирования до 10% промоторной области гена BAX была статистически значимо больше и составила 98% относительно группы сравнения, в которой доля таких людей не превышала 73% (р < 0,00001).

DNA methylation plays an important role in carcinogenesis; there are many studies that investigate the degree of methylation of the entire genome, gene promoters, and non-coding elements in cancer cells, but much less information about changes of the methylation patterns in blood cells and links with the development of malignant neoplasms (MN). This study aimed to investigate the degree of methylation of promoter regions of cell cycle control and apoptosis genes (BAX, MDM2, TP53, NFkB1) in peripheral blood cells of persons chronically exposed to radiation with MN developing latently. The study included 200 persons chronically exposed to radiation from the Techa River, contaminated with nuclear wastes dumped into it. The level of methylation was assessed by real-time PCR. The participants were divided into exposed and control groups; comparing them, we found that in the former, the distribution of exposed individuals with latent MN by the degree of methylation of promoter regions of BAX, MDM2 and NFkB1 genes was significantly different from that in the latter (p < 0.001; p < 0.001; p = 0.004, respectively). It was established that, compared to the control group, the share of the test group participants with subsequent MN who had up to 10% of the BAX gene promoter regions methylated was significantly higher, and amounted to 98%, while in the control group this figure did not exceed 73% (p < 0.00001).

хроническое радиационное воздействиеметилирование геновCpG-динуклеотидыканцерогенезрека Течаchronic radiation exposuregene methylationCpG dinucleotidescarcinogenesisthe Techa RiverReferencesSmith P, McGuffog L, Easton DF, Mann GJ, Pupo GM, Newman B, et al. A genome wide linkage search for breast cancer susceptibility genes. Genes Chromosomes Cancer. 2006; 45 (7): 646–55. DOI: 10.1002/gcc.20330.Smith P, McGuffog L, Easton DF, Mann GJ, Pupo GM, Newman B, et al. A genome wide linkage search for breast cancer susceptibility genes. Genes Chromosomes Cancer. 2006; 45 (7): 646–55. DOI: 10.1002/gcc.20330.Barnoud T, Parris JLD, Murphy ME. Common genetic variants in the TP53 pathway and their impact on cancer. J Mol Cell Biol. 2019; 11 (7): 578–85. DOI: 10.1093/jmcb/mjz052.Barnoud T, Parris JLD, Murphy ME. Common genetic variants in the TP53 pathway and their impact on cancer. J Mol Cell Biol. 2019; 11 (7): 578–85. DOI: 10.1093/jmcb/mjz052.Hall MJ, Bernhisel R, Hughes E, Larson K, Rosenthal ET, Singh NA, et al. Germline Pathogenic Variants in the Ataxia Telangiectasia Mutated (ATM) Gene are Associated with High and Moderate Risks for Multiple Cancers. Cancer Prev Res (Phila). 2021; 14 (4): 433–40. DOI: 10.1158/1940-6207.CAPR-20-0448.Hall MJ, Bernhisel R, Hughes E, Larson K, Rosenthal ET, Singh NA, et al. Germline Pathogenic Variants in the Ataxia Telangiectasia Mutated (ATM) Gene are Associated with High and Moderate Risks for Multiple Cancers. Cancer Prev Res (Phila). 2021; 14 (4): 433–40. DOI: 10.1158/1940-6207.CAPR-20-0448.Verma M, Rogers S, Divi RL, Schully SD, Nelson S, Joseph Su, et al. Epigenetic research in cancer epidemiology: trends, opportunities, and challenges. Cancer Epidemiol Biomarkers Prev. 2014; 23 (2): 223–33. DOI: 10.1158/1055-9965.EPI-13- 0573.Verma M, Rogers S, Divi RL, Schully SD, Nelson S, Joseph Su, et al. Epigenetic research in cancer epidemiology: trends, opportunities, and challenges. Cancer Epidemiol Biomarkers Prev. 2014; 23 (2): 223–33. DOI: 10.1158/1055-9965.EPI-13-0573.Esteller M, Corn PG, Baylin SB, Herman JG. A gene hypermethylation profile of human cancer. Cancer Research. 2001; 61 (8): 3225–9. PubMed PMID: 11309270.Esteller M, Corn PG, Baylin SB, Herman JG. A gene hypermethylation profile of human cancer. Cancer Research. 2001; 61 (8): 3225–9. PubMed PMID: 11309270.Belinsky SA. Gene-promoter hypermethylation as a biomarker in lung cancer. Nat Rev Cancer. 2004; 4 (9): 707–17. DOI: 10.1038/ nrc1432.Belinsky SA. Gene-promoter hypermethylation as a biomarker in lung cancer. Nat Rev Cancer. 2004; 4 (9): 707–17. DOI: 10.1038/ nrc1432.Park JY. Promoter hypermethylation in prostate cancer. Cancer Control. 2010; 17(4): 245–55. DOI: 10.1177/107327481001700405.Park JY. Promoter hypermethylation in prostate cancer. Cancer Control. 2010; 17(4): 245–55. DOI: 10.1177/107327481001700405.Suzuki K, Suzuki I, Leodolter A, Alonso S, Horiuchi S, Yamashita K, et al. Global DNA demethylation in gastrointestinal cancer is age dependent and precedes genomic damage. Cancer Cell. 2006; 9(3): 199–207. DOI: 10.1016/j.ccr.2006.02.016.Suzuki K, Suzuki I, Leodolter A, Alonso S, Horiuchi S, Yamashita K, et al. Global DNA demethylation in gastrointestinal cancer is age dependent and precedes genomic damage. Cancer Cell. 2006; 9(3): 199–207. DOI: 10.1016/j.ccr.2006.02.016.Ehrlich M. DNA hypomethylation in cancer cells. Epigenomics. 2009; 1 (2): 239–59. DOI: 10.2217/epi.09.33Ehrlich M. DNA hypomethylation in cancer cells. Epigenomics. 2009; 1 (2): 239–59. DOI: 10.2217/epi.09.33Wild CP, Scalbert A, Herceg Z. Measuring the exposome: a powerful basis for evaluating environmental exposures and cancer risk. Environ Mol Mutagen. 2013; 54 (7): 480–99. DOI: 10.1002/em.21777.Wild CP, Scalbert A, Herceg Z. Measuring the exposome: a powerful basis for evaluating environmental exposures and cancer risk. Environ Mol Mutagen. 2013; 54 (7): 480–99. DOI: 10.1002/ em.21777.Relton CL, Davey SG. Epigenetic epidemiology of common complex disease: prospects for prediction, prevention, and treatment. PLoS Med. 2010; 7 (10): e1000356. DOI: 10.1371/journal.pmed.1000356.Relton CL, Davey SG. Epigenetic epidemiology of common complex disease: prospects for prediction, prevention, and treatment. PLoS Med. 2010; 7 (10): e1000356. DOI: 10.1371/ journal.pmed.1000356.Suter CM, Martin DI, Ward RL. Germline epimutation of MLH1 in individuals with multiple cancers. Nat Genet. 2004; 36 (5): 497– 501. DOI: 10.1038/ng1342.Suter CM, Martin DI, Ward RL. Germline epimutation of MLH1 in individuals with multiple cancers. Nat Genet. 2004; 36 (5): 497–501. DOI: 10.1038/ng1342.Zheng Y, Joyce BT, Colicino E, Liu L, Zhang W, Dai Q, et al. Blood epigenetic age may predict cancer incidence and mortality. EBioMedicine. 2016; 5: 68–73. DOI: 10.1016/j.ebiom.2016.02.008.Zheng Y, Joyce BT, Colicino E, Liu L, Zhang W, Dai Q, et al. Blood epigenetic age may predict cancer incidence and mortality. EBioMedicine. 2016; 5: 68–73. DOI: 10.1016/j. ebiom.2016.02.008.Durso DF, Bacalini MG, Sala C, Pirazzini C, Marasco E, Bonafé M, et al. Acceleration of leukocytes' epigenetic age as an early tumor and sex-specific marker of breast and colorectal cancer. Oncotarget. 2017; 8 (14): 23237–45. DOI: 10.18632/oncotarget.15573.Durso DF, Bacalini MG, Sala C, Pirazzini C, Marasco E, Bonafé M, et al. Acceleration of leukocytes' epigenetic age as an early tumor and sex-specific marker of breast and colorectal cancer. Oncotarget. 2017; 8 (14): 23237–45. DOI: 10.18632/oncotarget.15573.Kresovich JK, Xu Z, O'Brien KM, Weinberg CR, Sandler DP, Taylor JA. Methylation-based biological age and breast cancer risk. J Natl Cancer Inst. 2019; 111 (10): 1051–8. DOI: 10.1093/jnci/djz020.Kresovich JK, Xu Z, O'Brien KM, Weinberg CR, Sandler DP, Taylor JA. Methylation-based biological age and breast cancer risk. J Natl Cancer Inst. 2019; 111 (10): 1051–8. DOI: 10.1093/jnci/djz020.Dugué PA, Bassett JK, Wong EM, Joo JE, Li S, Yu C. Biological aging measures based on blood DNA methylation and risk of cancer: a prospective study. JNCI Cancer Spectr. 2020; 5 (1): pkaa109. DOI: 10.1093/jncics/pkaa109.Dugué PA, Bassett JK, Wong EM, Joo JE, Li S, Yu C. Biological aging measures based on blood DNA methylation and risk of cancer: a prospective study. JNCI Cancer Spectr. 2020; 5 (1): pkaa109. DOI: 10.1093/jncics/pkaa109.Wang C, Ni W, Yao Y, Just A, Heiss J, Wei Y. DNA methylation- based biomarkers of age acceleration and all-cause death, myocardial infarction, stroke, and cancer in two cohorts: The NAS, and KORA F4. EBioMedicine. 2021; 63: 103151. DOI: 10.1016/j.ebiom.2020.103151.Wang C, Ni W, Yao Y, Just A, Heiss J, Wei Y. DNA methylation- based biomarkers of age acceleration and all-cause death, myocardial infarction, stroke, and cancer in two cohorts: The NAS, and KORA F4. EBioMedicine. 2021; 63: 103151. DOI: 10.1016/j.ebiom.2020.103151.Li X, Schöttker B, Holleczek B, Brenner H. Associations of DNA methylation algorithms of aging and cancer risk: Results from a prospective cohort study. EBioMedicine. 2022; 81: 104083. DOI: 10.1016/j.ebiom.2022.104083.Li X, Schöttker B, Holleczek B, Brenner H. Associations of DNA methylation algorithms of aging and cancer risk: Results from a prospective cohort study. EBioMedicine. 2022; 81: 104083. DOI: 10.1016/j.ebiom.2022.104083.Ennour-Idrissi K, Dragic D, Durocher F, Diorio C. Epigenome-wide DNA methylation and risk of breast cancer: a systematic review. BMC Cancer. 2020; 20 (1): 1048. DOI: 10.1186/s12885-020-07543-4.Ennour-Idrissi K, Dragic D, Durocher F, Diorio C. Epigenome-wide DNA methylation and risk of breast cancer: a systematic review. BMC Cancer. 2020; 20 (1): 1048. DOI: 10.1186/s12885-020- 07543-4.Дегтева М. О., Напье Б. А., Толстых Е. И., Шишкина Е. А., Бугров Н. Г., Крестинина Л. Ю. и др. Распределение индивидуальных доз в когорте людей, облученных в результате радиоактивного загрязнения реки Течи. Медицинская радиология и радиационная безопасность. 2019; 64 (3): 46–53. DOI: 10.12737/article_5cf2364cb49523.98590475.Degteva MO, Napier BA, Tolstykh EI, Shishkina EA, Bougrov NG, Krestinina LYu, et al. Individual dose distribution in cohort of people exposed as a result of radioactive contamination of the Techa River. Medical Radiology and Radiation Safety. 2019; 64 (3): 46–53. Russian.Никифоров В. С. Содержание матричной РНК генов, вовлеченных в клеточный гомеостаз человека, в отдаленные сроки после хронического облучения [диссертация]. Обнинск: 2021.Nikiforov VS. Soderzhanie matrichnoy RNK genov, vovlechennykh v kletochnyy gomeostaz cheloveka, v otdalennye sroki posle khronicheskogo oblucheniya [dissertation]. Obninsk; 2021. Russian.Blinova EA, Nikiforov VS, Kotikova AI, Yanishevskaya MA, Akleyev AV. Methylation Status of Apoptosis Genes and Intensity of Apoptotic Death of Peripheral Blood Lymphocytes in Persons Chronically Exposed to Radiation. Mol Biol (Mosk). 2022; 56 (6): 1072–82. DOI 10.1134/S002689332205003X.Blinova EA, Nikiforov VS, Kotikova AI, Yanishevskaya MA, Akleyev AV. Methylation Status of Apoptosis Genes and Intensity of Apoptotic Death of Peripheral Blood Lymphocytes in Persons Chronically Exposed to Radiation. Mol Biol (Mosk). 2022; 56 (6): 1072–82. DOI 10.1134/S002689332205003X.Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 1993; 74 (4): 609–19. DOI: 10.1016/0092-8674(93)90509-o.Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 1993; 74 (4): 609–19. DOI: 10.1016/0092-8674(93)90509-o.Gopisetty G, Ramachandran K, Singal R. DNA methylation and apoptosis. Mol Immunol. 2006; 43 (11): 1729–40. DOI: 10.1016/j. molimm.2005.11.010.Gopisetty G, Ramachandran K, Singal R. DNA methylation and apoptosis. Mol Immunol. 2006; 43 (11): 1729–40. DOI: 10.1016/j. molimm.2005.11.010.Alipour M, Zargar SJ, Safarian S, Fouladdel S, Azizi E, Jafargholizadeh N. The study of DNA methylation of BAX gene promoter in breast and colorectal carcinoma cell lines. Iran J Cancer Prev. 2013; 6 (2): 59–64.Alipour M, Zargar SJ, Safarian S, Fouladdel S, Azizi E, Jafargholizadeh N. The study of DNA methylation of BAX gene promoter in breast and colorectal carcinoma cell lines. Iran J Cancer Prev. 2013; 6 (2): 59–64.Блинова Е. А., Котикова А. И., Аклеев А. В. Интенсивность апоптоза лимфоцитов крови у облученных лиц с облигатными формами предраковых заболеваний. Бюллетень экспериментальной биологии и медицины. 2023; 176 (8): 233–6.Blinova EA, Kotikova AI, Akleev AV. The intensity of blood lymphocytes apoptosis in exposed individuals with obligate forms of precancerous conditions. Bulletin of Experimental Biology and Medicine. 2023; 176 (8): 233–6. Russian.Verma M, Rogers S, Divi RL, Schully SD, Nelson S, Joseph Su L, et al. Epigenetic research in cancer epidemiology: trends, opportunities, and challenges. Cancer Epidemiol Biomarkers Prev. 2014; 23 (2): 223–33. DOI: 10.1158/1055-9965.EPI-13-0573.Verma M, Rogers S, Divi RL, Schully SD, Nelson S, Joseph Su L, et al. Epigenetic research in cancer epidemiology: trends, opportunities, and challenges. Cancer Epidemiol Biomarkers Prev. 2014; 23 (2): 223–33. DOI: 10.1158/1055-9965.EPI-13-0573.Flanagan JM, Munoz-Alegre M, Henderson S, Tang T, Sun P, Johnson N, et al. Gene-body hypermethylation of ATM in peripheral blood DNA of bilateral breast cancer patients. Hum Mol Genet. 2009; 18 (7): 1332–42. DOI: 10.1093/hmg/ddp033.Flanagan JM, Munoz-Alegre M, Henderson S, Tang T, Sun P, Johnson N, et al. Gene-body hypermethylation of ATM in peripheral blood DNA of bilateral breast cancer patients. Hum Mol Genet. 2009; 18 (7): 1332–42. DOI: 10.1093/hmg/ddp033.

The authors declare that there are no conflicts of interest present.