Study of the intensity of free radical oxidation in mitochondria of endometrioid adenocarcinoma cells depending on the degree of tumor differentiation

Purpose of the study. To investigate redox-balance parameters in mitochondria of endometrioid adenocarcinoma (EA) cells in oncogynecological patients, as well as in mitochondria of Guerin carcinoma cells with intramural tumor localization in rats.

Materials and methods. The clinical part of the study included patients who underwent surgical treatment for EA (n = 42) and uterine fibroids (n = 14). Among patients with EA, 16 had well-differentiated tumors (G1), 12 had moderately differentiated tumors (G2), and 14 had poorly differentiated tumors (G3). The mean age of patients with EA was 60.8 ± 2.9 years, and with fibroids – 49.4 ± 2.5 years. None of the patients received neoadjuvant therapy. The experimental part of the study included biological material obtained from non-linear white female laboratory rats (n = 15) weighing 250 ± 25 g. Intramural growth of Guerin carcinoma was reproduced in these animals. In mitochondria isolated from EA, fibroid, and intact uterine tissues, concentrations of the following parameters were determined using standard ELISA methods: mitochondrial superoxide dismutase (SOD‑2), DNA and RNA oxidative modification products, malondialdehyde (MDA), diene conjugates (DC), and total protein (biuret method). Statistical analysis was performed using Statistica 10.0.

Results. In women, mitochondria of EA cells in G1 tumors demonstrated a markedly elevated content of MDA and DC – 2.3‑fold and 2.9‑fold higher, respectively, compared with values in mitochondria of intact uterine tissue. In G2 tumors, MDA levels were 3.2‑fold higher and DC levels 2.7‑fold higher than in intact mitochondria. In G3 tumors, the degree of DNA damage increased 1.6‑fold (p < 0.05), while MDA, DC, and SOD‑2 concentrations increased 2.4‑fold, 3.0‑fold, and 3.4‑fold, respectively, compared with intact values. In female rats, mitochondria isolated from tumor tissues with intramural Guerin carcinoma growth displayed changes similar in direction to the clinical results: DNA damage increased 1.6‑fold (p < 0.05), MDA 1.6‑fold (p < 0.05), DC 1.5‑fold (p < 0.05), and SOD‑2 2.4‑fold.

Conclusion. The oxidative stress and mitochondrial dysfunction identified in malignant cells of women indicate a differentiationdependent pathogenetic feature. Additionally, an identical direction of redox-balance alterations was demonstrated in the experimental subcellular model, supporting the translational relevance of the findings.

1. Lucas E, Carrick KS. Low grade endometrial endometrioid adenocarcinoma: A review and update with emphasis on morphologic variants, mimics, immunohistochemical and molecular features. Semin Diagn Pathol. 2022;39(3):159–175. https://doi.org/10.1053/j.semdp.2022.02.002

2. Kovalenko NV, Kit OI, Maksimov AYu, Demidova AA. Рossibilities of screening for cancer of the uterine body by the content of molecular markers in urine and vaginal-cervical secretion. Russian Clinical Laboratory Diagnostics. 2023;68(3):141–145. (In Russ.). https://doi.org/10.51620/0869-2084-2023-68-3-141-145

3. Žalytė E. Ferroptosis, metabolic rewiring, and endometrial cancer. Int J Mol Sci. 2023;25(1):75. https://doi.org/10.3390/ijms25010075

4. Tian H, Gao Z, Wang G, Li H, Zheng J. Estrogen potentiates reactive oxygen species (ROS) tolerance to initiate carcinogenesis and promote cancer malignant transformation. Tumour Biol. 2016 Jan;37(1):141–150. https://doi.org/10.1007/s13277-015-4370-6

5. Olowofolahan A, Fatunsin O, Olorunsogo O. Modulatory effect of ciprofloxacin, a broad spectrum antibacterial drug, on mPT pore using rat model with estradiol benzoate-induced endometrial hyperplasia. Naunyn Schmiedebergs Arch Pharmacol. 2024 May;397(5):3331–3341. https://doi.org/10.1007/s00210-023-02824-8

6. Tang S, Chen L. The recent advancements of ferroptosis of gynecological cancer. Cancer Cell Int. 2024 Oct 26;24(1):351. https://doi.org/10.1186/s12935-024-03537-5

7. Kit OI, Shikhlyarova AI, Frantsiyants EM, Neskubina IV, Kaplieva IV, Goncharova AS, et al. Processes of mitochondrial self-organization in experimental tumor growth with chronic neurogenic pain. Bulletin of Higher Educational Institutions. North Caucasus Region. Natural Sciences. 2019;2(202):97–105. (In Russ.).

8. Rospatent. A method for obtaining experimental malignant lung tumors. Sidorenko YuS, Frantsiyants EM, Komarova EF, Pogorelova YuA, Shikhlyarova AI. Patent for invention RU 2375758 C1, 12/10/2009. Application No. 2008133091/14 dated 08/11/20088. (In Russ.).

9. Kit OI, Frantsiyants EM, Shikhlyarova AI, Neskubina IV, Kaplieva IV, Cheryarina ND, et al. Biological effects of mitochondrial therapy: preventing development of myocardial infarction and blocking metastatic aggression of b16/f10 melanoma. Cardiometry. 2022;22:50–55. 10.18137/cardiometry.2022.22.5055

10. Dodokhova MA, Alkhusein-Kulyaginova MS, Safronenko AV, Kotieva IM, Shpakovsky DB, Milaeva ER. Effect of cisplatin and a hybrid organotin compound in low doses on the growth and metastasis of Lewis epidermoid carcinoma in an experiment. Experimental and Clinical Pharmacology 2021;84(8):32–35. (In Russ.). https://doi.org/10.30906/0869-2092-2021-84-8-32-35

11. Frantsiyants EM, Kaplieva IV, Bondovkina VA, Surikova EI, Neskubina IV, Trepitaki LK, et al. Modeling of multiple primary malignant tumors in experiment. South Russian Journal of Cancer. 2022;3(2):14–21. https://doi.org/10.37748/2686-9039-2022-3-2-2

12. Frantsiyants EM, Bandovkina VA, Kaplieva IV, Surikova EI, Neskubina IV, Pogorelova YuA, et al. Changes in pathophysiology of tumor growth and functional activity of the hypothalamic-pituitary-thyroid axis in rats of both sexes with the development of Guerin's carcinoma on the background of hypothyroidism. South Russian Journal of Cancer. 2022;3(4):26–39. https://doi.org/10.37748/2686-9039-2022-3-4-3

13. Rospatent. Method for creating an orthotopic endometrial cancer model. Frantsiyants EM, Shikhlyarova AI, Kaplieva IV, Bandovkina VA, Pogorelova YuA, Neskubina IV, et al. Patent for invention RU 2818464 C1, 05/02/2024. Application No. 2024103023 dated 02/07/2024. (In Russ.).

14. Frantsiyants EM, Shikhlyarova AI, Kaplieva IV, Bandovkina VA, Pogorelova YuA, Neskubina IV, et al. Establishment of an orthotopic model of endometrial cancer. Siberian Journal of Oncology. 2024;23(6):70–80. (In Russ.). https://doi.org/10.21294/1814-4861-2024-23-6-70-80

15. Egorova MV, Afanasev SA. Isolation of mitochondria from animal and human cells and tissues: Modern methodological techniques. Siberian Journal of Oncology. 2011;26(1–1):22–28. (In Russ.).

16. Gureev AP, Kokina AV, Syromyatnikov MYu, Popov VN. Optimization of methods for the mitochondria isolation from different mice tissues. Proceedings of Voronezh State University. Series: Chemistry. Biology. Pharmacy. 2015;4:61–65. (In Russ.).

17. Arutyunyan AV, Dubinina EE, Zybina NN. Methods for assessing free radical oxidation and the antioxidant system of the body. Methodological recommendations. St. Petersburg, 2000, 104 p. (In Russ.).

18. Sies H, Belousov VV, Chandel NS, Davies MJ, Jones DP, Mann GE, et al. Defining roles of specific reactive oxygen species (ROS) in cell biology and physiology. Nat Rev Mol Cell Biol. 2022 Jul;23(7):499–515. https://doi.org/10.1038/s41580-022-00456-z

19. Jomova K, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, Valko M. Several lines of antioxidant defense against oxidative stress: antioxidant enzymes, nanomaterials with multiple enzyme-mimicking activities, and low-molecular-weight antioxidants. Arch Toxicol. 2024 May;98(5):1323–1367. https://doi.org/10.1007/s00204-024-03696-4

20. Halliwell B. Understanding mechanisms of antioxidant action in health and disease. Nat Rev Mol Cell Biol. 2024 Jan;25(1):13–33. https://doi.org/10.1038/s41580-023-00645-4

21. Kim YS, Gupta Vallur P, Phaëton R, Mythreye K, Hempel N. Insights into the Dichotomous Regulation of SOD2 in Cancer. Antioxidants (Basel). 2017 Nov 3;6(4):86. https://doi.org/10.3390/antiox6040086

22. Zhou C, Lyu LH, Miao HK, Bahr T, Zhang QY, Liang T, et al. Redox regulation by SOD2 modulates colorectal cancer tumorigenesis through AMPK-mediated energy metabolism. Mol Carcinog. 2020 May;59(5):545–556. https://doi.org/10.1002/mc.23178 Erratum in: Mol Carcinog. 2023 Aug;62(8):1242–1243. https://doi.org/10.1002/mc.23595

23. Quirós I, Sáinz RM, Hevia D, García-Suárez O, Astudillo A, Rivas M, Mayo JC. Upregulation of manganese superoxide dismutase (SOD2) is a common pathway for neuroendocrine differentiation in prostate cancer cells. Int J Cancer. 2009 Oct 1;125(7):1497–1504. https://doi.org/10.1002/ijc.24501