The number of cancer stem cells in the tumor tissue and perifocal tissue of non-muscle invasive bladder cancer
https://doi.org/10.37748/2686-9039-2022-3-1-1
Abstract
Purpose of the study. Determine the content of cancer stem cells (CSCs) in the tumor tissue (TT) and perifocal tissues (PT) in muscle-non-invasive bladder cancer.
Materials and methods. We’ve examined fragments of TT and PT of 7 muscle-non-invasive bladder cancer (NMIBC) after surgical intervention – transurethral resection of the urinary bladder (TUR). In tissue samples that were used to obtain cell suspension of TT and PT using the BD Medimachine apparatus (BD, USA) was treated with monoclonal antibodies CD45-APCCy7, CD44-FITC, CD133-РЕ, CD24-PE (BD, USA) and were assessed on flow cytometer FacsCantoII (BD, USA). The percentage of cells with CSC phenotypic markers was determined in the analysis sample: CD45-CD44+CD24+, CD45-CD44+, CD45-CD24+, CD45-CD133+, CD45-CD44+CD133+. The presence of significant differences in the groups was evaluated using the STATISTICA 13 software package and the differences between the samples were considered significant at p < 0.05. The percentage of cells of the corresponding phenotype was calculated relative to the total number of cells. The percentage of cells with the corresponding phenotype was calculated relative to the total number of cells.
Results. The relative numbers of cells with CSC phenotypic markers, such as CD24, CD44, were 77 % and 58 % higher in TT than in PT: 18.3 ± 3.5 vs. 4.3 ± 2.1, p ≤ 0.044 and 15.5 ± 5.3 vs. 6.5 ± 0.8, p ≤ 0.043, respectively. The number of CD133+ cells was 83 % higher in PT compared to TT – 41.6 ± 12.1 vs. 22.7 ± 7.6, p ≤ 0.047.
Conclusion. The study of CSCs is a promising direction for the study of oncogenesis and can be used to assess the nature of the further development of relapse and / or progression of the disease, as well as various therapeutic approaches that are aimed at eliminating with CSC phenotypic markers and blocking the pathways leading to the emergence and maintenance of this cell population in patients with NMIBC.
About the Authors
L. I. BelyakovaRussian Federation
Lyubov I. Belyakova – PhD student
SPIN: 3382-8559, AuthorID: 1080471, ResearcherID: AAH-7729-2020
63 14 line str., Rostov-on-Don 344037, Russian Federation
A. N. Shevchenko
Russian Federation
Alexey N. Shevchenko – Dr. Sci. (Med.), Professor, Head of the Oncourology Department
SPIN: 2748-2638, AuthorID: 735424, ResearcherID: Y-5387-2018, Scopus Author ID: 57192283096
Rostov-on-Don
A. B. Sagakyants
Russian Federation
Aleksandr B. Sagakyants – Cand. Sci. (Biol.), associate Professor, Head of the Laboratory of Tumor Immunophenotyping
SPIN: 7272-1408, AuthorID: 426904, ResearcherID: M-8378-2019, Scopus Author ID: 24329773900
Rostov-on-Don
E. S. Bondarenko
Russian Federation
Elena S. Bondarenko – junior research fellow at the Laboratory of Tumor Immunophenotyping
SPIN: 3117-4040, AuthorID: 865798, Scopus Author ID: 57200132337
Rostov-on-Don
O. G. Shulgina
Russian Federation
Oksana G. Shulgina – junior research fellow at the Laboratory of Tumor Immunophenotyping
SPIN: 9668-3042, AuthorID: 886334
Rostov-on-Don
E. P. Ulyanova,
Russian Federation
Elena P. Ulyanova – junior research fellow at the Laboratory of Tumor Immunophenotyping
SPIN: 1243-9475, AuthorID: 759154, Scopus Author ID: 57203357998
Rostov-on-Don
E. V. Filatova
Russian Federation
Elena V. Filatova – Cand. Sci. (Med.), MD, oncologist, research fellow
SPIN: 7517-1549, AuthorID: 794870, Scopus Author ID: 5719228349
Rostov-on-Don
I. A. Khomutenko
Russian Federation
Irina A. Khomutenko – Cand. Sci. (Med.), senior research fellow
SPIN: 5401-5810, AuthorID: 735408
Rostov-on-Don
References
1. Kit OI, Franciyanc EM, Dimiriadi SN, Kaplieva IV, Trepitaki LK. Neoangiogenesis and fibrinolytic system biomarkers expression in the dynamics of experimental kidney ischemia in rats. Experimental and clinical urology. 2015;(1):20–23. (In Russ.).
2. Hakenberg OW (Chair), Compérat E, Minhas S, Necchi A, Protzel C, Watkin N. (Vice-chair) Guidelines Associate: Robinson R. EAU Guidelines on Penile Cancer. European Association of Urology, 2020. Available at: https://uroweb.org/guideline/penile-cancer/. Accessed: 17.01.2022.
3. Dzidzaria AG, Pavlov AYu, Gafanov RA, Fastovets SV, Kravtsov IB. Current issues in molecular diagnostics of bladder cancer. RMJ 2019;(2):56–60. (In Russ.).
4. Malignant neoplasms in Russia in 2020 (morbidity and mortality). Ed. by Kaprina AD, Starinsky VV, Shakhzadova AO, 2021, 252 p. Available at: https://oncology-association.ru/wp-content/uploads/2021/11/zis-2020-elektronnaya-versiya.pdf, Accessed: 17.01.2022. (In Russ.).
5. The state of oncological care to the population of Russia in 2019. Ed. by AD Kaprin, VV Starinsky, AO Shakhzadova. Moscow: P. A. Hertsen Moscow Oncology Research Institute – Branch of the National Medical Research Radiological Centre 2020, 239 p. (In Russ.).
6. Mikhaylenko DS, Sergienko SA, Zaborsky IN, Saflullin KN, Serebryany SA, Safronova NYu, et al. The role of molecular genetic alterations in sensitivity of the adjuvant intravesical therapy for non-muscle invasive bladder cancer. Cancer Urology. 2018;14(4):124–138. (In Russ.). https://doi.org/10.17650/1726-9776-2018-14-4-124-138
7. Osmanov YuI, Gaibov ZhA, Tursunov KhZ, Demyashkin GA, Barzak RI. Molecular characteristics of urotelial carcinomas of the urinary system. Crimea Journal of Experimental and Clinical Medicine 2019;9(2):76–82. (In Russ.).
8. Kit OI, Shevchenko AN, Komarova EF, Pakus DI, Maksimov AJu. Effect of conjugation matrix metalloproteinase genes polymorphism and their tissue inhibitors with the activity of extracellular proteolysis basement membrane components at early recurrence in patients with superficial bladder cancer. Ural Medical Journal 2015;(7(130)):73–78. (In Russ.).
9. Mikhaylenko DS, Perepechin DV, Efremov GD, Sivkov AV, Apolikhin OI. Detection of FGFR3 and PIK3CA mutations in DNA isolated from urine sediment of bladder cancer patients. Experimental and Clinical Urology. 2015;(4):38–41. (In Russ.).
10. Robertson AG, Kim J, Al-Ahmadie H, Bellmunt J, Guo G, Cherniack AD, et al. Comprehensive Molecular Characterization of Muscle-Invasive Bladder Cancer. Cell. 2017 Oct 19;171(3):540–556.e25. https://doi.org/10.1016/j.cell.2017.09.007
11. Segovia C, Martínez-Fernández M, Dueñas M, Rubio C, López-Calderón FF, Costa C, et al. Opposing roles of PIK3CA gene alterations to EZH2 signaling in non-muscle invasive bladder cancer. Oncotarget. 2017 Feb 7;8(6):10531–10542. https://doi.org/10.18632/oncotarget.14453
12. Sergienko SA, Mikhaylenko DS, Safronova NY, Efremov GD, Kaprin AD, Alekseev BYа. Somatic mutation profiling and functioning of intracellular signaling pathways at various stages of bladder cancer and their significance for targeted therapy. Experimental and Clinical Urology. 2020;(1):42–51. https://doi.org/10.29188/2222-8543-2020-12-1-42-51
13. Burger M, Catto JWF, Dalbagni G, Grossman HB, Herr H, Karakiewicz P, et al. Epidemiology and risk factors of urothelial bladder cancer. Eur Urol. 2013 Feb;63(2):234–241. https://doi.org/10.1016/j.eururo.2012.07.033
14. Belyakova LI, Shevchenko AN, Sagakyants AB, Filatova EV. Markers of bladder cancer: their role and prognostic significance (literature review). Cancer Urology. 2021;17(2):145–156. (In Russ.). https://doi.org/10.17650/17269776-2021-17-2-145-156
15. Puchinskaya MV. Cancer stem cell markers and their prognostic value. Arkhiv Patologii. 2016;78(2):47–54. (In Russ.). https://doi.org/10.17116/patol201678247-54
16. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. 1997 Jul;3(7):730–737. https://doi.org/10.1038/nm0797-730
17. Chan KS, Espinosa I, Chao M, Wong D, Ailles L, Diehn M, et al. Identification, molecular characterization, clinical prognosis, and therapeutic targeting of human bladder tumor-initiating cells. Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):14016–14021. https://doi.org/10.1073/pnas.0906549106
18. Yang Z, Li C, Fan Z, Liu H, Zhang X, Cai Z, et al. Single-cell Sequencing Reveals Variants in ARID1A, GPRC5A and MLL2 Driving Self-renewal of Human Bladder Cancer Stem Cells. Eur Urol. 2017 Jan;71(1):8–12. https://doi.org/10.1016/j.eururo.2016.06.025
19. Van Batavia J, Yamany T, Molotkov A, Dan H, Mansukhani M, Batourina E, et al. Bladder cancers arise from distinct urothelial sub-populations. Nat Cell Biol. 2014 Oct;16(10):982–991. https://doi.org/10.1038/ncb3038
20. Li Y, Lin K, Yang Z, Han N, Quan X, Guo X, et al. Bladder cancer stem cells: clonal origin and therapeutic perspectives. Oncotarget. 2017 Sep 12;8(39):66668–66679. https://doi.org/10.18632/oncotarget.19112
21. Wu C-T, Lin W-Y, Chang Y-H, Chen W-C, Chen M-F. Impact of CD44 expression on radiation response for bladder cancer. J Cancer. 2017;8(7):1137–1144. https://doi.org/10.7150/jca.18297
22. Yu X, Lin Y, Yan X, Tian Q, Li L, Lin EH. CD133, Stem Cells, and Cancer Stem Cells: Myth or Reality? Curr Colorectal Cancer Rep. 2011 Dec;7(4):253–259. https://doi.org/10.1007/s11888-011-0106-1
23. Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T, et al. CD133 expression is not restricted to stem cells, and both CD133+ and CD133- metastatic colon cancer cells initiate tumors. J Clin Invest. 2008 Jun;118(6):2111–2120. https://doi.org/10.1172/JCI34401
24. Huang P, Watanabe M, Kaku H, Ueki H, Noguchi H, Sugimoto M, et al. Cancer stem cell-like characteristics of a CD133+ subpopulation in the J82 human bladder cancer cell line. Mol Clin Oncol. 2013 Jan;1(1):180–184. https://doi.org/10.3892/mco.2012.29
25. Rola RM, Sammour SA-E, Shehab ElDin ZA, Salman MI, Omran TI. Expression of CD133 and CD24 and their different phenotypes in urinary bladder carcinoma. Cancer Manag Res. 2019;11:4677–4690. https://doi.org/10.2147/CMAR.S19834
26. Ortiz-Montero P, Liu-Bordes W-Y, Londoño-Vallejo A, Vernot J-P. CD24 expression and stem-associated features define tumor cell heterogeneity and tumorigenic capacities in a model of carcinogenesis. Cancer Manag Res. 2018;10:5767–5784. https://doi.org/10.2147/CMAR.S176654
Supplementary files
Review
For citations:
Belyakova L.I., Shevchenko A.N., Sagakyants A.B., Bondarenko E.S., Shulgina O.G., Ulyanova, E.P., Filatova E.V., Khomutenko I.A. The number of cancer stem cells in the tumor tissue and perifocal tissue of non-muscle invasive bladder cancer. South Russian Journal of Cancer. 2022;3(1):6-14. https://doi.org/10.37748/2686-9039-2022-3-1-1