• V.M. Mikhailenko R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology
  • E.A. Domina R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology
  • V.S. Ivankova National Cancer Institute, Ministry of Health of Ukraine
  • L.I. Makovetska R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology
  • O.A. Glavin R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology
  • T.V. Khrulenko National Cancer Institute, Ministry of Health of Ukraine




apoptosis, cervical cancer, chromosome aberration, DNA double strand breaks, oxidative metabolism, peripheral blood lymphocytes


Background: The combination of chemo- and radiotherapy used as main treatment of locally advanced cervical cancer (CC) may lead to side effects in healthy cells, which undermine the effectiveness of treatment and quality of life. The assessment of damage level in healthy radiosensitive cells from the tumor environment before the treatment is important in order to predict and prevent remote side effects of radiation. Aim: To study the oxidative metabolism and genetic disorders in peripheral blood lymphocytes (PBL) of primary CC patients in order to evaluate the possibilities of predicting radiation complications based on the molecular and biological properties of PBL. Materials and Methods: Peripheral blood samples were collected from 13 primary CC patients T1–4N0–1M0–1, and PBL were routinely isolated. The oxidative metabolism (mitochondrial trans-membrane potential, superoxide anion radical (О2•) generation, reactive oxygen species (ROS) production in PBL as well as the level of SH-groups in plasma and pro/antioxidant ratio in hemolysates were examined. The development of genetic instability was determined by estimation of DNA double-strand breaks (DNA-DSB), frequency and spectrum of chromosome aberrations and apoptosis. Results: The marked increase in the intensity of О2• generation in PBL (1.5-fold), depletion of SH-groups content (1.6-fold) and a shift in the pro-antioxidant balance (1.4-fold) towards its prooxidant component were observed in the blood of primary CC patients as compared to healthy individuals. These oxidative stress related events were accompanied by an increase in the level of DNA-DSB (2.1-fold), apoptosis (3.5-fold) and frequency of cells with chromosome aberrations (3.9-fold). On the contrary, significant decrease in mitochondrial trans-membrane potential (2.0-fold) and ROS generation in PBL (4.0-fold) were detected. Conclusion: Preliminary data indicate a violation of redox processes regulation, a shift in the pro-antioxidant balance towards its pro-oxidant component, accompanied by an increase in the level of DNA damage, development of genetic instability and apoptotic death of blood lymphocytes in primary CC patients.


Wild CP, Weiderpass E, Stewart BW, eds. World Cancer Report: Cancer research for cancer prevention. Lyon, France: International Agency for Research on Cancer; 2020. 611 p. https://digitallibrary.in.one.un.org/TempPdfFiles/5932_1.pdf. Accessed September 27, 2022.

Fedorenko ZP, Sumkina OV, Horokh YEL, et al. CANCER IN UKRAINE, 2020-2021: Morbidity, mortality, indicators activities of the oncology service. Bulletin of the National Cancer Registry Ukraine 2022; 23: 129 p. (in Ukrainian).

Lushnikova PA, Sukhikh ES, Izhevsky PV, et al. Modern methods of radiation therapy for cervical cancer. Creative surgery and oncology 2021; 11: 58–67. (in Russian). https://doi.org/10.24060/2076-3093-2021-11-1-58-677

NCCN clinical practice guidelines in oncology: cervical cancer. National Comprehensive Cancer Network, version I.2018 — October 25, 2017. https://oncolife.com.ua/doc/nccn/Cervical_Cancer.pdf. Accessed September 27, 2022.

Sarabhail T, Schaarschmidt BM, Wetter A, et al. Comparison of 18F-FDG PET/MRI and MRI for pre-therapeutic tumor staging of patients with primary cancer of the uterine cervix. Eur J Nucl Med Mol 2018; 45: 67–76. https://doi.org/10.1186/s40644-020-00372-5

Martinelli F, Signorelli M, Bogani G, et al. Is aortic lymphadenectomy indicated in locally advanced cervical cancer after neoadjuvant chemotherapy followed by radical surgery? A retrospective study on 261 women. Eur J Surg Oncol 2016; 42: 1512–8. https://doi.org/10.1016/j.ejso.2016.06.004

Meirow D, Nugent D. The effects of radiotherapy and chemotherapy on female reproduction. Hum Reprod Update 2001; 7: 535–43. https://doi.org/10.1093/humupd/7.6.535

Ye S, Yang J, Cao D, et al. A systematic review of quality of life and sexual function of patients with cervical cancer after treatment. Int J Gynecol Cancer 2014; 24: 1146–57. https://doi.org/10.1097/IGC.0000000000000207

Mkrtchian LS, Ivanov SA, Kulieva GZK, et al. Quality of life of patients with cervical cancer after radiotherapy and chemoradiotherapy. Radiation Risk 2020; 29: 120–8 (in Russian). https://doi.org/10.21870/0131-3878-2020-29-1-120-128

Han X, Yang Q, Zhang J, et al. Correlation between changes in the number of peripheral blood lymphocytes and survival rate in patients with cervical cancer after radio-chemotherapy. Cancer Radiother 2021; 25: 72–6. https://doi.org/10.1016/j.canrad.2020.08.045

Gavrilescu MM, Hutanu I, Ioanid N, et al. Clinical value of hematological biomarkers in uterine cervical cancer. Chirurgia (Bucur) 2016; 111: 493–9. https://doi.org/10.21614/chirurgia.111.6.493

Jeong MH, Kim H, Kim TH, et al. Prognostic significance of pretreatment lymphocyte percentage and age at diagnosis in patients with locally advanced cervical cancer treated with definite radiotherapy. Obstet Gynecol Sci 2019; 62: 35–45. https://doi.org/10.5468/ogs.2019.62.1.35

Armon-Omer A, Neuman H, Sharabi-Nov A, et al. Mitochondrial activity is impaired in lymphocytes of MS patients in correlation with disease severity. Mult Scler Relat Disord 2020; 41: 102025. https://doi.org/10.1016/j.msard.2020.102025

Takabayashi A, Kanai M, Kawai Y, et al. Change in mitochondrial membrane potential in peripheral blood lymphocytes, especially in natural killer cells, is a possible marker for surgical stress on the immune system. World J Surg 2003; 27: 659–65. https://doi.org/10.1007/s00268-003-6926-7

Georgescu SR, Mitran CI, Mitran MI, et al. New insights in the pathogenesis of HPV infection and the associated carcinogenic processes: the role of chronic inflammation and oxidative stress. J Immunol Res 2018; 2018: 5315816. https://doi.org/10.1155/2018/5315816

Zahra K, Patel S, Dey T, et al. A study of oxidative stress in cervical cancer — an institutional study. Biochem Biophys Rep 2021; 25: 100881. https://doi.org/10.1016/j.bbrep.2020.100881

Everett S.A., Wardman P. Perthiols as antioxidants: radical-scavenging and pro-oxidative mechanisms. Methods Enzymol 1995; 251: 55–69. https://doi.org/10.1016/0076-6879(95)51110-5

Scott D. Chromosomal radiosensitivity, cancer predisposition and response to radiotherapy. Strahlenther Onkol 2000; 176: 229–34. https://doi.org/10.1007/s000660050005

Ivankova VS, Domina EA, Khrulenko TV, et al. Effects of brachytherapy on cytogenetic parameters and oxidative status in peripheral blood lymphocytes of gynecologic cancer patients. Exp Oncol 2021; 43: 242–6. https://doi.org/10.32471/exp-oncology.2312-8852.vol-43-no-3.16514

Product Information Histopaque®-1077 Hybri-Max™ (H8889). https://www.sigmaaldrich.com/ content/dam/ sigma-aldrich/docs/ Sigma/ Product_Information_Sheet/ 2/h8889pis.pdf. Accessed October 17, 2019.

Bokunyaeva NI, Zolotnitskaya RP. Handbook of Clinical Laboratory Research Methods. M: Medicine, 1975. 384 p. (in Russian).

Sivandzade F, Bhalerao A, Cucullo L. Analysis of the mitochondrial membrane potential using the cationic JC-1 de as a sensitive fluorescent probe. Bio Protoc 2019; 9: e3128. https://doi.org/10.21769/BioProtoc.3128. http://www.bio-protocol.org/e3128. Accessed September 27, 2022.

MitoPT® JC-1 Assay Manual. ImmunoChemistry Technologies, LLC. #F18-911-8-G, 8 p. https://cdn.shopify.com/s/files/1/0549/4210/5739/files/911-924-mitopt-jc-1-assay-product-manual.pdf. Accessed September 27, 2022.

Glavin OA, Domina EA, Mikhailenko VM, et al. Metformin as a modifier of the oxidative state of peripheral blood and the viability of human lymphocytes under the influence of ionizing radiation. Oncologiya 2020; 22: 84–91 (in Ukrainian). https://doi.org/10.32471/oncology.2663-7928.t-22-1-2020-g.8855

Liochev SI, Fridovich I. Lucigenin (bis-N-methylacridinium) as a mediator of superoxide anion production. Arch Biochem Biophys 1997; 337: 115–20. https://doi.org/10.1006/abbi.1997.9766

Druzhyna МО, Makovetska LI, Glavin OA, et al. The free-radical processes in peripheral blood of patients with benign breast disease. Oncologiya 2018; 78: 250–4. (in Ukrainian).

Yao K, Wu W, Wang KJ, et al. Electromagnetic noise inhibits radiofrequency radiation-induced DNA damage and reactive oxygen species increase in human lens epithelial cells. Mol Vis 2008; 14: 964–9.

Tarpey MM, Wink DA, Grisham MB. Methods for detection of reactive metabolites of oxygen and nitrogen: in vitro and in vivo considerations. Am J Physiol Regul Integr Comp Physiol 2004; 286: R431–44. https://doi.org/10.1152/ajpregu.00361.2003

Serkiz YaI, Druzhina NA, Khriyenko AP, et al. Blood Chemiluminescence Under Radiation Exposure. K: Naukova Dumka, 1989. 176 p. (in Russian).

Olive PL, Banáth JP. The comet assay: a method to measure DNA damage in individual cells. Nat Protoc 2006; 1: 23–9. https://doi.org/10.1038/nprot.2006.5

Mikhailenko VM, Muzalov II. Exogenous nitric oxide potentiate DNA damage and alter DNA repair in cells exposed to ionizing radiation. Exp Oncol 2013; 35: 318–24.

Collins AR. Investigating oxidative DNA damage and its repair using the comet assay. Mutat Res 2009; 681: 24–32. https://doi.org/10.1016/j.mrrev.2007.10.002

Cytogenetic Dosimetry: Applications in Preparedness for and Response to Radiation Emergencies. Vienna: IAEA, 2011. 232 p.

Riccardi C, Nicoletti I. Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 2006; 1: 1458–61. https://doi.org/10.1038/nprot.2006.238

Hu ML. Measurement of protein thiol groups and glutathione in plasma. Methods Enzymol 1994; 233: 380–5. https://doi.org/10.1016/s0076-6879(94)33044-1

Miwa S, Kashyap S, Chini E, et al. Mitochondrial dysfunction in cell senescence and aging. J Clin Invest 2022; 132: e158447. https://doi.org/10.1172/JCI158447

Song B, Li T, Chen S, et al. Correlations between MTP and ROS levels of peripheral blood lymphocytes and readmission in patients with chronic heart failure. Heart Lung Circ 2016; 25: 296–302. https://doi.org/10.1016/j.hlc.2015.09.004

Armon-Omer A, Neuman H, Sharabi-Nov A, et al. Mitochondrial activity is impaired in lymphocytes of MS patients in correlation with disease severity. Mult Scler Relat Disord 2020; 41: 102025. https://doi.org/10.1016/j.msard.2020.102025. https://doi.org/10.1016/j.msard.2020.102025.

Anaya-Eugenio GD, Tan CY, Rakotondraibe LH, et al. Tumor suppressor p53 independent apoptosis in HT-29 cells by auransterol from Penicillium aurantiacobrunneum. Biomed Pharmacother 2020; 127: 110124. https://doi.org/10.1016/j.biopha.2020.110124

Zhu P, Luo W, Qian J, et al. GSH/ROS Dual-responsive supramolecular nanoparticles based on pillar[6]arene and betulinic acid prodrug for chemo-chemodynamic combination therapy. Molecules 2021; 26: 5900. https://doi.org/10.3390/molecules26195900

Wang Y, Yin S, Zhou Y, et al. Dual-function of baicalin in nsPEFs-treated hepatocytes and hepatocellular carcinoma cells for different death pathway and mitochondrial response. Int J Med Sci 2019; 16: 1271–82. https://doi.org/10.7150/ijms.34876

Perl A, Gergely PJr, Banki K, et al. Mitochondrial dysfunction in T cells of patients with systemic lupus erythematosus. Int Rev Immunol 2004; 23: 293–313. https://doi.org/10.1080/08830180490452576

Lee DS, Schrader A, Warchol M, et al. Cisplatin exposure acutely disrupts mitochondrial bioenergetics in the zebrafish lateral-line organ. Hear Res 2022; 4: 108513. https://doi.org/10.1016/j.heares.2022.108513

Banjerdpongchai R, Wudtiwai B, Sringarm K. Cytotoxic and apoptotic-inducing effects of purple rice extracts and chemotherapeutic drugs on human cancer cell lines. Asian Pac J Cancer Prev 2014; 14: 6541–8. https://doi.org/10.7314/apjcp.2013.14.11.6541

Goroshinskaya I; Popova N; Menshenina A, et al. Free radical processes in the blood of patients with cervical cancer receiving various postoperative treatment modalities. Int J Gynecol Cancer 2019; 29: A218. https://doi.org/10.1136/ijgc-2019-ESGO.362

Shah S, Kalal BS. Oxidative stress in cervical cancer and its response to chemoradiation. Turk J Obstet Gynecol 2019; 16: 124–8. https://doi.org/10.4274/tjod.galenos.2019.19577

Chernikova NV, Goroshinskaya IA, Frantsiyants EM, et al. Intensity of free-radical reactions in metastasizing cervical cancer. Meeting abstract, 2021 ASCO annual meeting. J Clin Oncol 2021; 39: e17508. https://doi.org/10.1200/JCO.2021.39.15_suppl.e17508

Monaghan P, Metcalfe NB, Torres R. Oxidative stress as a mediator of life history trade-offs: mechanisms, measurements and interpretation. Ecol Lett 2009; 12: 75-92. https://doi.org/10.1111/j.1461-0248.2008.01258.x

Ott M, Gogvadze V, Orrenius S, et al. Mitochondria, oxidative stress and cell death. Apoptosis 2007; 12: 913-22. https://doi.org/10.1007/s10495-007-0756-2

Domina E. Expediency on using radiomitigators in radiation therapy of cancer patients. Journal of science. Lyon. France 2020. 1: 7–11. http://www.joslyon.com/wp-content/uploads/2020/08/Lyon_10_1.pdf. Accessed September 27, 2022.

Chekhun VF, Domina EA. Can SARS-CoV-2 change individual radiation sensitivity of the patients recovered from COVID-19? (Experimental and theoretical background). Exp Oncol 2021; 43: 277–80. https://doi.org/10.32471/exp-oncology.2312-8852.vol-43-no-3.16554

Domina Е. Possible effects of the exposure to ionizing radiation on the patients recovered from COVID-19. ScienceRise: Biol Sci 2022; 1: 4-7. doi: http://doi.org/10.15587/2519-8025.2022.254881




How to Cite

Mikhailenko, V., Domina, E., Ivankova, V., Makovetska, L., Glavin, O., & Khrulenko, T. (2023). FEATURES OF OXIDATIVE METABOLISM AND GENETIC DISORDERS IN PERIPHERAL BLOOD LYMPHOCYTES OF PATIENTS WITH PRIMARY CERVICAL CANCER. Experimental Oncology, 44(3), 227–233. https://doi.org/10.32471/exp-oncology.2312-8852.vol-44-no-3.18486



Original contributions