Mitochondrial apoptosis factors in colorectal cancer tissue

Purpose of the study. To examine the influence of specific factors on the apoptosis process in the mitochondria of cells located in various regions of the colon, with a focus on patients of both sexes.

Patients and methods. The study included results obtained from 132 patients with T2–3N0M0 colon cancer, comprising 52 women and 80 men. Mitochondria were isolated from human intestinal and tumor tissue cells using differential centrifugation. The concentration of cytochrome C (ng/mg protein), AIF (pg/mg protein), Bcl-2 (pg/mg protein), and calcium (mM/mg protein) were determined.

Results. In men, the calcium level in the mitochondria of both rectal tumor tissue and left colon tumor tissue was reduced relative  to the corresponding tissue along the resection line, while the levels of Bcl-2, cytochrome C, and AIF were, on the contrary, in creased. At the same time, the calcium and Bcl-2 levels in the mitochondria of the right colon tumor tissue did not differ statistically  significantly from the corresponding values in the right colon tissue along the resection line, while the levels of cytochrome C and  AIF were increased by 2.0 and 3.1 times. The calcium level was observed to be reduced by a factor of 1.5 in the mitochondria of rectal tumor tissue cells in women, in comparison to the indicator in the rectal tissue along the resection line (p < 0.05). The levels of Bcl-2, Cytochrome C, and AIF were increased by 1.5, 2.9, and 2.1 times, respectively. In the mitochondria of tumor tissue in the  left half of the colon of women, the calcium level was reduced relative to the indicator in the tissue of the left half of the colon  along the resection line by 1.6 times (p < 0.05), and AIF was statistically significantly increased by 2.7 times. In women, the levels  of cytochrome C and AIF in the mitochondria of tumor tissue cells in the right half of the colon were 2.0 and 1.7 times higher,  respectively, than the corresponding indicators in the tissue of the right half of the colon along the resection line (p < 0.05).

Conclusion. A comprehensive analysis of the studied indicators suggests that apoptosis is suppressed in all regions of the large intestine in both men and women. Conversely, the processes of respiration and mitochondrial energy production appear to be enhanced.

1. Patel SG, Karlitz JJ, Yen T, Lieu CH, Boland CR. The rising tide of early-onset colorectal cancer: a comprehensive review of epidemiology, clinical features, biology, risk factors, prevention, and early detection. Lancet Gastroenterol Hepatol. 2022;7(3):262–274. https://doi.org/10.1016/s2468-1253(21)00426-x

2. Kit OI, Dzhenkova EA, Mirzoyan EA, Gevorkyan YuA, Sagakyants AB, Timoshkina NN, et al. Molecular genetic classification of colorectal cancer subtypes: current state of the problem. South Russian Journal of Cancer. 2021;2(2):50–56. https://doi.org/10.37748/2686-9039-2021-2-2-6

3. Abancens M, Bustos V, Harvey H, McBryan J, Harvey BJ. Sexual Dimorphism in Colon Cancer. Front Oncol. 2020 Dec 9;10:607909. https://doi.org/10.3389/fonc.2020.607909

4. Wu Z, Xiao C, Long J, Huang W, You F, Li X. Mitochondrial dynamics and colorectal cancer biology: mechanisms and potential targets. Cell Comm Sig. 2024;22(1):91. https://doi.org/10.1186/s12964-024-01490-4

5. Zhang L, Yu J. Role of apoptosis in colon cancer biology, therapy, and prevention. Curr Colorectal Cancer Rep. 2013 Dec;9(4):10. https://doi.org/10.1007/s11888-013-0188-z

6. Kit OI, Shikhlyarova AI, Frantsiyants EM, Neskubina IV, Kaplieva IV, Goncharova AS, et al. Processes of mitochondrial self-organiza tion in experimental tumor growth with chronic neurogenic pain. Bulletin Of Higher Education Institutes. North Caucasus Region. Natural Sciences. 2019;2(202):97–105. (In Russ.). https://doi.org/10.23683/0321-3005-2019-2-97-105

7. Ramachandran A, Madesh M, Balasubramanian KA. Apoptosis in the intestinal epithelium: its relevance in normal and pathophysiological conditions. J Gastroenterol Hepatol. 2000 Feb;15(2):109–120. https://doi.org/10.1046/j.1440-1746.2000.02059.x

8. Bock FJ, Tait SWG. Mitochondria as multifaceted regulators of cell death. Nat Rev Mol Cell Biol. 2020;21(2):85–100. https://doi.org/10.1038/s41580-019-0173-8

9. Egorova MV, Afanasyev SA. Isolation of mitochondria from cells and tissues of animals and human: modern methodical approaches. Siberian Medical Journal. 2011;26(1–1):22–28. (In Russ.).

10. 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.).

11. Carew JS, Huang P. Mitochondrial defects in cancer. Mol Cancer. 2002;1:9. https://doi.org/10.1186/1476-4598-1-9

12. Lagadinou ED, Sach A, Callahan K, Rossi RM, Nearing SJ, Minhajuddin M, et al. BCL-2 inhibition targets oxidative phosphorylation and selectively kills quiescent human leukemia stem cells. Cell Stem Cells. 2013;12(3):329–341. https://doi.org/10.1016/j.stem.2012.12.013

13. Panov A, Orynbayeva Z. Bioenergetic and antiapoptotic properties of mitochondria from cultured human prostate cancer cell lines PC-3, DU145 and LNCaP. PLoS One. 2013;8(8):e72078. https://doi.org/10.1371/journal.pone.0072078

14. Zhang BB, Wang DG, Guo FF, Xuan C. Mitochondrial membrane potential and reactive oxygen species in cancer stem cells. FamCancer. 2015;14(1):19–23. https://doi.org/10.1007/s10689-014-9757-9

15. Sullivan LB, Chandel NS. Mitochondrial reactive oxygen species and cancer. Metabol Cancer. 2014;2:17. https://doi.org/10.1186/2049-3002-2-17

16. Corbet C, Pinto A, Martherus R, Santiago de Jesus JP, Polet F, Feron O. Acidosis drives the reprogramming of fatty acid metabolism in cancer cells through changes in mitochondrial and histone acetylation. Cell Metab. 2016;24(2):311–323. https://doi.org/10.1016/j.cmet.2016.07.003

17. Moon DO. The role of calcium in orchestrating apoptosis in cancer: a mitochondrial perspective. Int J Mol Sci. 2023;24(10):8982. https://doi.org/10.3390/ijms24108982

18. Pinton P, Giorgi C, Siviero R, Zecchini E, Rizzuto R. Calcium and apoptosis: ER-mitochondria Ca2+ transfer in the control of apoptosis. Oncogene. 2008 Oct 27;27(50):6407–6418. https://doi.org/10.1038/onc.2008.308

19. Patergnani S, Danese A, Bouhamida E, Aguiari G, Previati M, Pinton P, Giorgi C. Various Aspects of Calcium Signaling in the Regulation of Apoptosis, Autophagy, Cell Proliferation, and Cancer. Int J Mol Sci. 2020 Nov 6;21(21):8323. https://doi.org/10.3390/ijms21218323

20. Eckenrode EF, Yang J, Velmurugan GV, Foskett JK, White C. Apoptosis protection by Mcl-1 and Bcl-2 modulation of inositol 1,4,5-trisphosphate receptor-dependent Ca2+ signaling. J Biol Chem. 2010 Apr 30;285(18):13678–13684. https://doi.org/10.1074/jbc.M109.096040

21. Chang MJ, Zhong F, Lavik AR, Parys JB, Berridge MJ, Distelhorst CW. Feedback regulation mediated by Bcl-2 and DARPP-32 regulates inositol 1,4,5-trisphosphate receptor phosphorylation and promotes cell survival. Proc Natl Acad Sci USA. 2014 Jan 21;111(3):1186– 1191. https://doi.org/10.1073/pnas.1323098111

22. Vervloessem T, Kerkhofs M, La Rovere RM, Sneyers F, Parys JB, Bultynck G. Bcl-2 inhibitors as anti-cancer therapeutics: The impact of and on calcium signaling. Cell Calcium. 2018 Mar;70:102–116. https://doi.org/016/j.ceca.2017.05.014

23. Andreu-Fernandez V, Sancho M, Genoves A, Lucendo E, Todt F, Lauterwasser J, et al. The Bax transmembrane domain interacts with Bcl-2 pro-survival proteins in biological membranes. Proc Natl Acad Sci USA. 2017;114(2):310–315. https://doi.org/1073/pnas.1612322114

24. Ramesh P, Medema JP. BCL-2 family deregulation in colorectal cancer: potential for BH3 mimetics in therapy. Apoptosis. 2020;25(5 6):305–320. https://doi.org/10.1007/s10495-020-01601-9

25. Cui J, Placzek WJ. Post-Transcriptional regulation of anti-apoptotic BCL2 family members. Int J Mol Sci. 2018;19(1):308. https://doi.org/10.3390/ijms19010308

26. Lindner AU, Salvucci M, Morgan C, Monsefi N, Resler AJ, Cremona M, et al. BCL-2 system analysis identifies high-risk colorectal cancer patients. Gut. 2017;66(12):2141–2148. https://doi.org/10.1136/gutjnl-2016-312287

27. Xu L, Xie Q, Qi L, Wang C, Xu N, Liu W, et al. Bcl-2 overexpression reduces cisplatin cytotoxicity by decreasing ER-mitochondrial Ca2+ signaling in SKOV3 cells. Oncol Rep. 2018;39(3):985–992. https://doi.org/10.3892/or.2017.6164

28. Singh R, Letai A, Sarosiek K. Regulation of apoptosis in health and disease: the balancing effect of BCL-2 family proteins. Nat Rev Mol Cell Biology. 2019;20(1):175–193. https://doi.org/10.1038/s41580-018-0089-8

29. Rana R, Huirem RS, Kant R, Chauhan K, Sharma S, Yashavarddhan MH, et al. Cytochrome C as a potential clinical marker for diag nosis and treatment of glioma. Front Oncol. 2022;12:960787. https://doi.org/10.3389/fonc.2022.960787

30. Morse PT, Arroum T, Wan J, Pham L, Vaishnav A, Bell J, et al. Phosphorylations and Acetylations of Cytochrome c Control Mitochondrial Respiration, Mitochondrial Membrane Potential, Energy, ROS, and Apoptosis. Cells. 2024;13(6):493. https://doi.org/10.3390/cells13060493

31. Culpage HA, Wang J, Morse PT, Zurek MP, Turner AA, Khobeir A, et al. Cytochrome c phosphorylation: control of electron flow in the mitochondrial electron transport chain and apoptosis. Int. J. Biochem. Cell Biol. 2020;121:105704. https://doi.org/10.1016/j.biocel.2020.105704

32. Cheng TS, Hong K, Eiki YW, Yuan S, Eiki KW. Near-atomic structure of the active human apoptosome. eLife. 2016;5:e17755. https://doi.org/10.7554/eLife.17755

33. Kalkavan H, Chen MJ, Crawford JC, Quarato G, Fitzgerald P, Tait SWG, Goding CR, Green DR. Sublethal cytochrome c release generates drug-tolerant persister cells. Cell. 2022 Sep 1;185(18):3356-3374.e22. https://doi.org/1016/j.cell.2022.07.025

34. González-Arzola C, Díaz-Quintana A, Bernardo-García N, Martínez-Fábregas J, Rivero-Rodríguez F, Casado-Combreras MA, et al. Nuclear-translocated mitochondrial cytochrome c releases the nucleophosmin-sequestered tumor suppressor ARF by altering nucleolar fluid-phase partitioning. Nat Struct Mol Biol. 2022 Oct;29(10):1024–1036. https://doi.org/10.1038/s41594-022-00842

35. Kalpage HA, Bazylianska V, Recanati MA, Fite A, Liu J, Wan J, et al. Tissue-specific regulation of cytochrome c by post-translational modifications: respiration, the mitochondrial membrane potential, ROS, and apoptosis. FASEB J. 2019;33(2):1540–1553. https://doi.org/10.1096/fj.201801417R

36. Novo N, Romero-Tamayo S, Marcuello C, Boneta S, Blasco-Machin I, Velazquez-Campoy A, et al. A platform protein for degradosome assembly: apoptosis-inducing factor as an efficient nuclease involved in chromatinolysis. PNAS Nexus. 2022;2(2):pgac312. https://doi.org/10.1093/pnasnexus/pgac312

37. Norberg E, Orrenius S, Zhivotovsky B. Mitochondrial regulation of cell death: processing of apoptosis-inducing factor (AIF). Bio chem Biophys Res Commun. 2010;396(1):95–100. https://doi.org/10.1016/j.bbrc.2010.02.163

38. Sorrentino L, Calogero AM, Pandini V, Vanoni MA, Sevrioukova IF, Aliverti A. Key role of the adenylate moiety and integrity of the adenylate-binding site for the NAD(+)/H binding to mitochondrial apoptosis-inducing factor. Biochemistry. 2015;54(47):6996–7009. https://doi.org/10.1021/acs.biochem.5b00898

39. Sorrentino L, Cossu F, Milani M, Aliverti A, Mastrangelo E. Structural bases of the altered catalytic properties of a pathogenic variant of apoptosis inducing factor. Biochem Biophys Res Commun. 2017;490(3):1011–1017. https://doi.org/10.1016/j.bbrc.2017.06.156

40. Brosey CA, Ho C, Long WZ, Singh S, Burnett K, Hura GL, et al. Defining NADH-Driven allostery regulating apoptosis-inducing factor. Structure. 2016;24(12):2067–2079. https://doi.org/10.1016/j.str.2016.09.012

41. Novo N, Ferreira P, Medina M. The apoptosis-inducing factor family: Moonlighting proteins in the crosstalk between mitochondria and nuclei. IUBMB Life. 2021;73(3):568–581. https://doi.org/10.1002/iub.2390

42. Dixon-Murray E, Nedara K, Mojtahedi N, Tokatlidis K. The Mia40/CHCHD4 oxidative folding system: regulation of redox processes and signaling in the mitochondrial intermembrane space. Antioxidants (Basel). 2021;10(4):592. https://doi.org/10.3390/antiox10040592

43. Bano D, Prehn JHM. Apoptosis-Inducing Factor (AIF) in Physiology and Disease: The Tale of a Repented Natural Born Killer. EBio Medicine. 2018 Apr;30:29–37. https://doi.org/10.1016/j.ebiom.2018.03.016

44. Wischhof L, Scifo E, Ehninger D, Bano D. AIFM1 beyond cell death: An overview of this OXPHOS-inducing factor in mitochondrial diseases. EBioMedicine. 2022 Sep;83:104231. 10.1016/j.ebiom.2022.104231. https://doi.org/10.1016/j.ebiom.2022.104231. Nat Struct Mol Biol. 2022 Oct;29(10):1024–1036. https://doi.org/10.1038/s41594-022-00842