vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJGB-22-08vavilov-3255Research ArticleГЕНЕТИКА ЧЕЛОВЕКАHUMAN GENETICSТранскрипционная активность генов репарации, апоптоза и клеточного цикла (TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3) у хронически облученных людей с различной интенсивностью апоптоза лимфоцитов периферической кровиTranscriptional activity of repair, apoptosis and cell cycle genes (TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3) in chronically exposed persons with different intensity of apoptosis of peripheral blood lymphocyteshttps://orcid.org/0000-0002-6685-1823НикифоровВ. С.NikiforovV. S.

Челябинск

 Chelyabinsk

nikiforovx@mail.ruhttps://orcid.org/0000-0002-2567-7945БлиноваЕ. А.BlinovaE. A.

Челябинск

 Chelyabinsk

https://orcid.org/0000-0002-1695-1340КотиковаА. И.KotikovaA. I.

Челябинск

 Chelyabinsk

https://orcid.org/0000-0003-2583-5808АклеевА. 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 России; Челябинский государственный университетРоссияUrals Research Center for Radiation Medicine; Chelyabinsk State UniversityRussian Federation2022010320222615058Copyright © Никифоров В.С., Блинова Е.А., Котикова А.И., Аклеев А.В., 20222022Никифоров В.С., Блинова Е.А., Котикова А.И., Аклеев А.В.Nikiforov V.S., Blinova E.A., Kotikova A.I., Akleyev A.V.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/3255

Исследовали транскрипционную активность генов, вовлеченных в поддержание генетического гомеостаза клетки (репарации, клеточного цикла и апоптоза: TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3), у лиц, подвергшихся хроническому радиационному облучению, с повышенной интенсивностью раннего, позднего апоптоза и некроза лимфоцитов периферической крови. Объектом изучения служили мононуклеарные клетки периферической крови, полученные от 132 жителей прибрежных сел реки Течи, подвергшихся хроническому облучению. Доза облучения красного костного мозга составляла 426.4±48.2 мГр (1.3–2930.0 мГр), доза облучения тимуса и периферических органов иммунной системы – 58.9±7.9 мГр (0.1–489.0 мГр). Исследование проводили в отдаленные сроки (более 60 лет с начала облучения), возраст людей на время проведения обследования был 68±0.6 года (55–86 лет). Анализ апоптотической и некротической гибели лимфоцитов периферической крови основывался на наличии на поверхности мембраны клеток фосфолипида фосфатидилсерина, а также ее проницаемости для интеркалирующего ДНК-красителя. Оценку относительного содержания мРНК генов репарации, клеточного цикла и апоптоза проводили с использованием полимеразной цепной реакции в реальном времени. В группе хронически облученных людей с повышенной интенсивностью раннего апоптоза отмечено увеличение относительного содержания мРНК гена PADI4 (p = 0.006). Для хронически облученных людей с повышенной интенсивностью позднего апоптоза зафиксирована модуляция относительного содержания мРНК генов TP53 (p = 0.013) и BCL-2 (p = 0.021). В отдаленные сроки у хронически облученных людей с повышенной интенсивностью некроза лимфоцитов периферической крови отмечен статистически значимый рост транскрипционной активности гена TP53 (p = 0.015). Установлено, что у облученных людей с повышенной интенсивностью апоптоза регистрируются в первую очередь изменения со стороны транскрипционной активности апоптотических генов, что согласуется с существующими представлениями об активации программированной гибели клеток.

Transcriptional activity of genes involved in maintaining genetic homeostasis (genes for repair, cell cycle and apoptosis: TP53, MDM2, ATM, BAX, BCL-2, CDKN1A, OGG1, XPC, PADI4, MAPK8, NF-KB1, STAT3, GATA3) was studied in chronically exposed persons with an increased intensity of early and late stages of apoptosis and necrosis of peripheral blood lymphocytes. The object of this study was peripheral blood mononuclear cells obtained from 132 chronically exposed residents of the Techa riverside villages. The mean accumulated dose to red bone marrow was 426.4±48.2 mGy (1.3–2930.0 mGy), to thymus and peripheral immune organs, 58.9±7.9 mGy (0.1–489.0 mGy). The study was performed more than 60 years after the onset of exposure, the average age of exposed persons was 68±0.6 years (55–86 years). The study of apoptotic and necrotic death of peripheral blood lymphocytes was based on the presence of phosphatidylserine on the cell membrane surface, as well as on its permeability for DNA-intercalating dye. Evaluation of the relative content of mRNA genes for repair, cell cycle, and apoptosis was carried out using real-time PCR. An increased relative content of PADI4 gene mRNA was registered in the group of chronically exposed persons with the increased intensity of early apoptosis (p = 0.006). Modulation of the relative content of mRNA of the TP53 (p = 0.013) and BCL-2 (p = 0.021) genes was detected in the group of chronically exposed individuals with the increased intensity of the late stage of apoptosis. A statistically significant increase in the transcriptional activity of the TP53 gene was observed in the group of chronically exposed persons with the increased intensity of peripheral blood lymphocyte necrosis in the long-term period (p = 0.015). In the course of the study it was noted that exposed people with increased intensity of apoptosis, first of all, demonstrate changes in the transcriptional activity of apoptotic genes. These data are consistent with current views on the activation of programmed cell death.

мРНКапоптознекрозлимфоцитыхроническое облучениеmRNAapoptosisnecrosislymphocyteschronic exposureReferencesAkleyev A.V. (Ed.) Consequences of Radioactive Contamination of the Techa River. Chelyabinsk: Kniga Publ., 2016. (in Russian)Akleyev A.V. (Ed.) Consequences of Radioactive Contamination of the Techa River. Chelyabinsk: Kniga Publ., 2016. (in Russian)Azimian H., Bahreyni-Toossi M.T., Rezaei A.R., Rafatpanah H., Hamzehloei T., Fardid H. Up-regulation of Bcl-2 expression in cultured human lymphocytes after exposure to low doses of gamma radiation. J. Med. Phys. 2015;40(1):38-44. DOI 10.4103/09716203.152249.Azimian H., Bahreyni-Toossi M.T., Rezaei A.R., Rafatpanah H., Hamzehloei T., Fardid H. Up-regulation of Bcl-2 expression in cultured human lymphocytes after exposure to low doses of gamma radiation. J. Med. Phys. 2015;40(1):38-44. DOI 10.4103/09716203.152249.Baryshnikov A.Yu., Shishkin Yu.V. Immunological Problems of Apoptosis. Moscow, 2002. (in Russian)Baryshnikov A.Yu., Shishkin Yu.V. Immunological Problems of Apoptosis. Moscow, 2002. (in Russian)Bevers E.M., Williamson P.L. Getting to the outer leaflet: physiology of phosphatidylserine exposure at the plasma membrane. Physiol. Rev. 2016;96(2):605-645. DOI 10.1152/physrev.00020.2015.Bevers E.M., Williamson P.L. Getting to the outer leaflet: physiology of phosphatidylserine exposure at the plasma membrane. Physiol. Rev. 2016;96(2):605-645. DOI 10.1152/physrev.00020.2015.Blinova E.A., Kotikova A.I., Yanishevskaya М.А., Akleyev A.V. Apoptosis of lymphocytes and polymorphisms of apoptosis regulation genes in individuals exposed to chronic radiation exposure. Meditsinskaya Radiologiya i Radiatsionnaya Bezopasnost’ = Medical Radiology and Radiation Safety. 2020a;65(4):36-42. DOI 10.12737/1024-6177-2020-65-4-36-42. (in Russian)Blinova E.A., Kotikova A.I., Yanishevskaya М.А., Akleyev A.V. Apoptosis of lymphocytes and polymorphisms of apoptosis regulation genes in individuals exposed to chronic radiation exposure. Meditsinskaya Radiologiya i Radiatsionnaya Bezopasnost’ = Medical Radiology and Radiation Safety. 2020a;65(4):36-42. DOI 10.12737/1024-6177-2020-65-4-36-42. (in Russian)Blinova E.A., Nikiforov V.S., Yanishevskaya M.A., Akleyev A.А. Single nucleotide polymorphism and expression of genes for immune competent cell proliferation and differentiation in radiationexposed individuals. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2020b;24(4):399-406. DOI 10.18699/VJ20.632. (in Russian)Blinova E.A., Nikiforov V.S., Yanishevskaya M.A., Akleyev A.А. Single nucleotide polymorphism and expression of genes for immune competent cell proliferation and differentiation in radiationexposed individuals. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2020b;24(4):399-406. DOI 10.18699/VJ20.632. (in Russian)Chipuk J.E. Dissecting p53-dependent apoptosis. Cell Death Differ. 2006;13(6):994-1002. DOI 10.1038/sj.cdd.4401908.Chipuk J.E. Dissecting p53-dependent apoptosis. Cell Death Differ. 2006;13(6):994-1002. DOI 10.1038/sj.cdd.4401908.Franco S., Alt F.W., Manis J.P. Pathways that suppress programmed DNA breaks from progressing to chromosomal breaks and translocations. DNA Repair. (Amst.). 2006;5(9-10):1030-1041. DOI 10.1016/j.dnarep.2006.05.024.Franco S., Alt F.W., Manis J.P. Pathways that suppress programmed DNA breaks from progressing to chromosomal breaks and translocations. DNA Repair. (Amst.). 2006;5(9-10):1030-1041. DOI 10.1016/j.dnarep.2006.05.024.Guimaraes D.P., Hainaut P. TP53: a key gene in human cancer. Biochimie. 2002;84(1):83-93. DOI 10.1016/s0300-9084(01)01356-6.Guimaraes D.P., Hainaut P. TP53: a key gene in human cancer. Biochimie. 2002;84(1):83-93. DOI 10.1016/s0300-9084(01)01356-6.Hahn W.C., Weinberg R.A. Modelling the molecular circuitry of cancer. Nat. Rev. Cancer. 2002;2(5):331-341. DOI 10.1038/nrc795.Hahn W.C., Weinberg R.A. Modelling the molecular circuitry of cancer. Nat. Rev. Cancer. 2002;2(5):331-341. DOI 10.1038/nrc795.Kabacik S., Mackay A., Tamber N., Manning G., Finnon P., Paillier F., Ashworth A., Bouffler S., Badie C. Gene expression following ionising radiation: identification of biomarkers for dose estimation and prediction of individual response. Int. J. Radiat. Biol. 2011;87(2):115-129. DOI 10.3109/09553002.2010.519424.Kabacik S., Mackay A., Tamber N., Manning G., Finnon P., Paillier F., Ashworth A., Bouffler S., Badie C. Gene expression following ionising radiation: identification of biomarkers for dose estimation and prediction of individual response. Int. J. Radiat. Biol. 2011;87(2):115-129. DOI 10.3109/09553002.2010.519424.Kheifets L.B., Abalakin V.A. Separation of human blood corpuscles in the verografin-ficoll density gradient. Laboratornoye Delo = Laboratory Science. 1973;10:579-581. (in Russian)Kheifets L.B., Abalakin V.A. Separation of human blood corpuscles in the verografin-ficoll density gradient. Laboratornoye Delo = Laboratory Science. 1973;10:579-581. (in Russian)Kvatcheva Yu.E. Human bone marrow repair processes and cell populations in acute radiation injury: morphological study. Radiatsionnaya Biologiya. Radioekologiya = Radiological Biology. Radioecology. 2000;40(1):5-9. (in Russian)Kvatcheva Yu.E. Human bone marrow repair processes and cell populations in acute radiation injury: morphological study. Radiatsionnaya Biologiya. Radioekologiya = Radiological Biology. Radioecology. 2000;40(1):5-9. (in Russian)Miyashita T., Krajewski S., Krajewska M., Wang H.G., Lin H.K., Liebermann D.A., Hoffman B., Reed J.C. Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene. 1994;9(6):1799-1805.Miyashita T., Krajewski S., Krajewska M., Wang H.G., Lin H.K., Liebermann D.A., Hoffman B., Reed J.C. Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene. 1994;9(6):1799-1805.Nikiforov V.S., Blinova E.A., Akleyev A.V. The transcriptional activity of cell cycle and apoptosis genes in chronically exposed people with an increased frequency of TCR mutant lymphocytes. Radiatsiya i Risk = Radiation and Risk. 2020;29(2):89-100. DOI 10.21870/0131-3878-2020-29-2-89-100. (in Russian)Nikiforov V.S., Blinova E.A., Akleyev A.V. The transcriptional activity of cell cycle and apoptosis genes in chronically exposed people with an increased frequency of TCR mutant lymphocytes. Radiatsiya i Risk = Radiation and Risk. 2020;29(2):89-100. DOI 10.21870/0131-3878-2020-29-2-89-100. (in Russian)Puthalakath H., Villunger A., O’Reilly L.A., Beaumont J.G., Coultas L., Cheney R.E., Huang D.C., Strasser A. BMF: a proapoptotic BH3-only protein regulated by interaction with the myosin V actin motor complex, activated by anoikis. Science. 2001;293(5536): 1829-1832. DOI 10.1126/science.1062257.Puthalakath H., Villunger A., O’Reilly L.A., Beaumont J.G., Coultas L., Cheney R.E., Huang D.C., Strasser A. BMF: a proapoptotic BH3-only protein regulated by interaction with the myosin V actin motor complex, activated by anoikis. Science. 2001;293(5536): 1829-1832. DOI 10.1126/science.1062257.Rogers G.E., Harding H.W., Llewellyn-Smith I.J. The origin of citrulline-containing proteins in the hair follicle and the chemical nature of trichohyalin, an intracellular precursor. Biochim. Biophys. Acta. 1977;495(1):159-175. DOI 10.1016/0005-2795(77)90250-1.Rogers G.E., Harding H.W., Llewellyn-Smith I.J. The origin of citrulline-containing proteins in the hair follicle and the chemical nature of trichohyalin, an intracellular precursor. Biochim. Biophys. Acta. 1977;495(1):159-175. DOI 10.1016/0005-2795(77)90250-1.Tanikawa C., Espinosa M., Suzuki A., Masuda K., Yamamoto K., Tsuchiya E., Ueda K., Daigo Y., Nakamura Y., Matsuda K. Regulation of histone modification and chromatin structure by the p53-PADI4 pathway. Nat. Commun. 2012;3:676. DOI 10.1038/ ncomms1676.Tanikawa C., Espinosa M., Suzuki A., Masuda K., Yamamoto K., Tsuchiya E., Ueda K., Daigo Y., Nakamura Y., Matsuda K. Regulation of histone modification and chromatin structure by the p53-PADI4 pathway. Nat. Commun. 2012;3:676. DOI 10.1038/ ncomms1676.Verheij M., Bartelink H. Radiation-induced apoptosis. Cell Tissue Res. 2000;301(1):133-142. DOI 10.1007/s004410000188.Verheij M., Bartelink H. Radiation-induced apoptosis. Cell Tissue Res. 2000;301(1):133-142. DOI 10.1007/s004410000188.Zeegers D., Venkatesan S., Koh S.W., Low G.K., Srivastava P., Sundaram N., Sethu S., Banerjee B., Jayapal M., Belyakov O., Baskar R., Balajee A.S., Hande M.P. Biomarkers of ionizing radiation exposure: A multiparametric approach. Genome Integr. 2017;8:6. DOI 10.4103/2041-9414.198911.Zeegers D., Venkatesan S., Koh S.W., Low G.K., Srivastava P., Sundaram N., Sethu S., Banerjee B., Jayapal M., Belyakov O., Baskar R., Balajee A.S., Hande M.P. Biomarkers of ionizing radiation exposure: A multiparametric approach. Genome Integr. 2017;8:6. DOI 10.4103/2041-9414.198911.

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