Apoptosis-Inducing Factor (AIF) content in tumor cell mitochondria from colorectal cancer patients

Purpose of the study. To investigate the level of protein AIF in the mitochondria of tumor cells and visually unchanged tissues of the colon in male and female patients with colorectal cancer.

Materials and methods. The study included results, obtained from 132 patients with stage T2–3N0M0 colon cancer, comprising 52 women and 80 men. Mitochondria were isolated from human colon and tumor tissue cells using differential centrifugation in a high-speed refrigerated centrifuge. The concentration of protein AIF (pg/mg protein) in mitochondria was determined using ELISA «Human AIF Elisa Kit» (Cloud-CloneCorp., China).

Results. It was established that in males, the AIF level in the mitochondria of rectal, sigmoid colon and ascending colon tumor cells was 2.4 times, 1.9 times (p < 0.05) and 3.1 times higher, respectively, than in the mitochondria of the corresponding tissues not affected by the tumor. In the mitochondria of the intestinal tissue not affected by the tumor, significant differences in the AIF content were observed, with levels varying depending on the anatomical location. In the sigmoid colon, the level of this factor was found to be 1.9 (p < 0.05) and 2.6 times higher than in the rectum and ascending colon, respectively. Concurrently, no notable discrepancies in the AIF concentration within the mitochondria of conditionally unimpaired tissues were observed in the female subjects. The AIF content was found to be higher in the mitochondria of tumor cells in women than in conditionally intact tissues. Specifically, it was observed to be 2.1 times higher in rectal tumors, 4.4 times higher in sigmoid colon tumors and 1.7 times (p < 0.05) higher in ascending colon tumors. Significant discrepancies in the AIF content between men and women, as well as between the mitochondria of tumor sample cells, were identified. In the rectal and ascending colon tumor, the AIF level in women was found to be markedly elevated in comparison to men, exhibiting a ratio of 1.3 (p < 0.05) and 2.4, respectively.

Conclusion. In patients with colorectal cancer, the content of AIF in tumor mitochondria is observed to increase. This can be considered to represent stimulation mechanism of tumor proliferative activity due to its NADH/NADPH oxidase function, which promotes the survival of malignant cells.

1. Jiang C., Zhou Q., Yi K., Yuan Y., Xie X. Colorectal cancer initiation: understanding early-stage disease for intervention. Cancer Lett. 2024589, 216831. doi: 10.1016/j.canlet.2024.216831.

2. Seo H.W., No H., Cheon H.J., Kim J.K. Sappanchalcone, a flavonoid isolated from Caesalpinia sappan L., induces caspase-dependent and AIF-dependent apoptosis in human colon cancer cells. Chem Biol Interact. 2020;327:109185. doi: 10.1016/j.cbi.2020.109185.

3. Hba S., Ghaddar S., Wahnou H., Pinon A., El Kebbaj R., Pouget C., Sol V., Liagre B., Oudghiri M., Limami Y. Natural chalcones and derivatives in colon cancer: pre-clinical challenges and the promise of chalcone-based nanoparticles. Pharmaceutics. 2023;15(12):2718. doi: 10.3390/pharmaceutics15122718.

4. Jiang C., Zhou Q., Yi K., Yuan Y., Xie X. Colorectal cancer initiation: understanding early-stage disease for intervention. Cancer Lett. 2024;589:216831. doi: 10.1016/j.canlet.2024.216831.

5. Abedizadeh R., Majidi F., Khorasani H.R., Abedi H., Sabour D. Colorectal cancer: a comprehensive review of carcinogenesis, diagnosis, and novel strategies for classified treatments. Cancer Metastasis Rev. 2024;43(2):729-753. doi: 10.1007/s10555-023-10158-3.

6. Zhang Y., Yan H., Wei Y., Wei X. Decoding mitochondria's role in immunity and cancer therapy. Biochim Biophys Acta Rev Cancer. 2024;1879(4):189107. doi: 10.1016/j.bbcan.2024.189107.

7. Prasad N., Sharma J.R., Yadav U.C.S. Induction of growth cessation by acacetin via β-catenin pathway and apoptosis by apoptosis inducing factor activation in colorectal carcinoma cells. Mol Biol Rep. 2020;47(2):987-1001. doi: 10.1007/s11033-019-05191-x.

8. Lin C., Chen X., Qiu H., Li B., Guo M. The Activation of the Tumor Suppressor Protein p53 by Acriflavine Leads to Mitochondrial Dysfunction and Improves the Radiosensitivity of Colon Cancer Cells. J Immunol Res. 2022;2022:1328542. doi: 10.1155/2022/1328542.

9. Bano D., Prehn J.H.M. Apoptosis-inducing factor (AIF) in physiology and disease: the tale of a repented natural born killer. EBioMedicine. 2018;30:29-37. doi: 10.1016/j.ebiom.2018.03.016.

10. Shen S.M., Guo M., Xiong Z., Yu Y., Zhao X.Y., Zhang F.F., Chen G.Q. AIF inhibits tumor metastasis by protecting PTEN from oxidation. EMBO Rep. 2015;16(11):1563-80. doi: 10.15252/embr.201540536.

11. Sevrioukova I.F. Apoptosis-inducing factor: structure, function, and redox regulation. Antioxid Redox Signal. 2011;14(12):2545-79. doi: 10.1089/ars.2010.3445.

12. Kit O.I., Frantsiyants E.M., Neskubina I.V., Cheryarina N.D., Shikhlyarova A.I., Przhedetskiy Y.V., Pozdnyakova V., Surikova E.I., Kaplieva I.V., Bandovkina V.A. Influence of standard and stimulated growth of b16/f10 melanoma on AIF levels in mitochondria in cells of the heart and other somatic organs in female mice Cardiometry. 2021;18:113-120. doi: 10.18137/cardiometry.2021.18.113120.

13. Egorova M.V., Afanas`ev S.A. Isolation of mitochondria from animal and human cells and tissues: Modern methodological techniques. Siberian Medical Journal. 2011;26(1-1):22-28. (in Russ.).

14. Gureev A.P., Kokina A.V., Sy`romyatnikova M.Yu., Popov V.N. Optimization of methods for isolating mitochondria from different mouse tissues. Bulletin of VSU, series: chemistry, biology, pharmacy. 2015;4:61-65. (in Russ.).

15. Lkhagvasuren K., Kim J.K. Ziyuglycoside II induces caspases-dependent and caspases-independent apoptosis in human colon cancer cells. Toxicol In Vitro. 2019;59:255-262. doi: 10.1016/j.tiv.2019.04.028.

16. Zong L., Liang Z. Apoptosis-inducing factor: a mitochondrial protein associated with metabolic diseases-a narrative review. Cardiovasc Diagn Ther. 2023;13(3):609-622. doi: 10.21037/cdt-23-123.

17. Lewis E.M., Wilkinson A.S., Jackson J.S., Mehra R., Varambally S., Chinnaiyan A.M., Wilkinson J.C. The enzymatic activity of apoptosis-inducing factor supports energy metabolism benefiting the growth and invasiveness of advanced prostate cancer cells. J Biol Chem. 2012;287(52):43862-75. doi: 10.1074/jbc.M112.407650.

18. Ademoni-Elisha L., Nakdimon I., Shteinfer A., Prezma T., Arif T., Arbel N., Melkov A., Zelichov O., Levi I., Shoshan-Barmatz V., et al. Novel biomarker proteins in chronic lymphocytic leukemia: Impact on diagnosis, prognosis and treatment. PLoS One. 2016;11(4):e0148500. doi: 10.1371/journal.pone.0148500.

19. Shoshan-Barmatz V., Arif T., Shteinfer-Kuzmine A. Apoptotic proteins with non-apoptotic activity: expression and function in cancer. Apoptosis. 2023;28(5-6):730-753. doi: 10.1007/s10495-023-01835-3.

20. Xing Y., Li Y., Hu B., Han F., Zhao X., Zhang H., Li Y., Li D., Li J., Jin F., Li F. PAK5-mediated AIF phosphorylation inhibits its nuclear translocation and promotes breast cancer tumorigenesis. Int J Biol Sci. 2021;17(5):1315-1327. doi: 10.7150/ijbs.58102.

21. Kadam A., Mehta D., Jubin T. et al. Apoptosis inducing factor: Cellular protective function in Dictyostelium discoideum. Biochim Biophys Acta Bioenerg. 2020;1861:148158. doi: 10.1016/j.bbabio.2020.148158.

22. Lin S.C., Chu P.Y., Liao WT. et al. Glycyrrhizic acid induces human MDA-MB-231 breast cancer cell death and autophagy via the ROS-mitochondrial pathway. Oncol Rep. 2018;39:703–10. doi: 10.3892/or.2017.6123.

23. Aburatani T., Inokuchi M., Takagi Y. et al. High expression of P21-activated kinase 5 protein is associated with poor survival in gastric cancer. Oncol Lett. 2017;14:404–10. 10.3892/ol.2017.6115.

24. Li Y.K., Zou J., Ye D.M. et al. Human p21-activated kinase 5 (PAK5) expression and potential mechanisms in relevant cancers: Basic and clinical perspectives for molecular cancer therapeutics. Life Sci. 2020;241:117113. doi: 10.1016/j.lfs.2019.117113.

25. Xing Y., Li Y., Hu B., Han F., Zhao X., Zhang H., Li Y., Li D., Li J., Jin F., Li F. PAK5-mediated AIF phosphorylation inhibits its nuclear translocation and promotes breast cancer tumorigenesis. Int J Biol Sci. 2021;17(5):1315-1327. doi: 10.7150/ijbs.58102.

26. Shoshan-Barmatz V., Arif T., Shteinfer-Kuzmine A. Apoptotic proteins with non-apoptotic activity: expression and function in cancer. Apoptosis. 2023;28(5-6):730-753. doi: 10.1007/s10495-023-01835-3.