EXPRESSION OF CD44 AND CD24 MARKERS IN BIOPSY SAMPLES OF TRIPLE NEGATIVE BREAST CANCER PATIENTS BEFORE TREATMENT

The role of the expression of CD44 and CD24 in breast cancer (BC) has been explored in many laboratories around the world to identify predictive markers of tumor aggressiveness and patient’s response to anticancer therapy. These proteins participate in the process of tumor growth, metastasis and formation of cancer stem cells (CSCs). The study of CD44 and CD24 expression in triple negative (TN) BC, which is the most aggressive breast cancer subtype, is of particular interest.
The aim of this study was to determine the relationship between the expression of CD44 and CD24 markers in biopsy samples of TNBC patients before treatment and clinical/ morphological characteristics of the tumors.
Material and Methods. The study group included 67 patients with stage I–IV TNBC. Flow cytometry was used to determine the proportion of cells with CSC immunophenotype (CD44+/CD24^-/low) in biopsy samples from the primary tumor of 65 patients and lymph nodes of 6 patients. In addition, the proportion of cells with all possible combinations of expression of these surface proteins was estimated.
Results. Cells with CSC immunophenotype were detected in all patients with a wide individual variability of CSC proportion from 0.4 % to 77.0 % (median – 10.9 %). There were no differences in the proportion of CSCs in the primary tumor and lymph nodes. No statistically significant correlation between the proportion of CSCs in the primary tumor and the clinical/morphological parameters, including tumor size and differentiation grade, evidence of regional or distant metastases, tumor, size of the fraction of proliferating cells estimated by Ki67 expression, was found in either single or multivariate analysis. There was also no association of the above parameters (except Ki67) with immunophenotypes. A high proportion of Ki67-positive cells in the primary tumor was associated with the CD44-CD24-phenotype. Conclusion. The expression of CD44 and CD24 in biopsy samples of TNBC before treatment did not correlate with the clinical and morphological characteristics of the tumors, excepting Ki67 expression.

triple negative breast cancer, cancer stem cells, CD44, CD24, flow cytometry

1. Honeth G., Bendahl P.O., Ringnér M., Saal L.H., Gruvberger-Saal S.K., Lövgren K., Grabau D., Fernö M., Borg A., Hegardt C. The CD44+/ CD24- phenotype is enriched in basal-like breast tumors. Breast Cancer Res. 2008; 10(3): R53. doi: 10.1186/bcr2108.

2. Ma F., Li H., Wang H., Shi X., Fan Y., Ding X., Lin C., Zhan Q., Qian H., Xu B. Enriched CD44(+)/CD24(-) population drives the aggressive phenotypes presented in triple-negative breast cancer (TNBC). Cancer Lett. 2014 Oct 28; 353(2): 153–9. doi: 10.1016/j.canlet.2014.06.022.

3. Shima H., Yamada A., Ishikawa T., Endo I. Are breast cancer stem cells the key to resolving clinical issues in breast cancer therapy? Gland Surg. 2017 Feb; 6(1): 82–88. doi: 10.21037/gs.2016.08.03.

4. Wang H., Wang L., Song Y., Wang S., Huang X., Xuan Q., Kang X., Zhang Q. CD44+/CD24- phenotype predicts a poor prognosis in triplenegative breast cancer. Oncol Lett. 2017 Nov; 14(5): 5890–5898. doi: 10.3892/ol.2017.6959.

5. Idowu M.O., Kmieciak M., Dumur C., Burton R.S., Grimes M.M., Powers C.N., Manjili M.H. CD44(+)/CD24(-/low) cancer stem/progenitor cells are more abundant in triple-negative invasive breast carcinoma phenotype and are associated with poor outcome. Hum Pathol. 2012 Mar; 43(3): 364–73. doi: 10.1016/j.humpath.2011.05.005.

6. Perrone G., Gaeta L.M., Zagami M., Nasorri F., Coppola R., Borzomati D., Bartolozzi F., Altomare V., Trodella L., Tonini G., Santini D., Cavani A., Muda A.O. In situ identification of CD44+/CD24- cancer cells in primary human breast carcinomas. PLoS One. 2012; 7(9): e43110. doi: 10.1371/journal.pone.0043110.

7. Camerlingo R., Ferraro G.A., De Francesco F., Romano M., Nicoletti G., Di Bonito M., Rinaldo M., D'Andrea F., Pirozzi G. The role of CD44+/CD24-/low biomarker for screening, diagnosis and monitoring of breast cancer. Oncol Rep. 2014 Mar; 31(3): 1127–32. doi: 10.3892/or.2013.2943.

8. Makki J., Myint O., Wynn A.A., Samsudin A.T., John D.V. Expression distribution of cancer stem cells, epithelial to mesenchymal transition, and telomerase activity in breast cancer and their association with clinicopathologic characteristics. Clin Med Insights Pathol. 2015; 8: 1–16. doi: 10.4137/CPath.S19615.

9. Abraham B.K., Fritz P., McClellan M., Hauptvogel P., Athelogou M., Brauch H. Prevalence of CD44+/CD24-/low cells in breast cancer may not be associated with clinical outcome but may favor distant metastasis. Clin Cancer Res. 2005 Feb 1; 11(3): 1154–9.

10. Tiezzi D.G., Valejo F.A., Marana H.R., Carrara H.H., Benevides L., Antonio H.M., Sicchieri R.D., Milanezi C.M., Silva J.S., de Andrade J.M. CD44+/CD24- cells and lymph node metastasis in stage I and II invasive ductal carcinoma of the breast. Med Oncol. 2012; 29(3): 1479–85. doi: 10.1007/s12032-011-0014-x.

11. Kai M., Onishi H., Souzaki M., Tanaka H., Kubo M., Tanaka M., Katano M. Semi-quantitative evaluation of CD44(+) /CD24(-) tumor cell distribution in breast cancer tissue using a newly developed fluorescence immunohistochemical staining method. Cancer Sci. 2011; 102(12): 2132–8. doi: 10.1111/j.1349-7006.2011.02063.x.

12. Guler G., Balci S., Costinean S., Ussakli C.H., Irkkan C., Suren D., Sari E., Altundag K., Ozisik Y., Jones S., Bacher J., Shapiro C.L., Huebner K. Stem cell-related markers in primary breast cancers and associated metastatic lesions. Mod Pathol. 2012 Jul; 25(7): 949–55. doi: 10.1038/modpathol.2012.37.

13. Zhong Y., Shen S., Zhou Y., Mao F., Guan J., Lin Y., Xu Y., Sun Q. ALDH1 is a better clinical indicator for relapse of invasive ductal breast cancer than the CD44+/CD24- phenotype. Med Oncol. 2014 Mar; 31(3): 864. doi: 10.1007/s12032-014-0864-0.

14. Liu C., Luo Y., Liu X., Lu P., Zhao Z. Clinical implications of CD44+/CD24- tumor cell ratio in breast cancer. Cancer Biother Radiopharm. 2012 Jun; 27(5): 324–8. doi: 10.1089/cbr.2011.1155.

15. Mylona E., Giannopoulou I., Fasomytakis E., Nomikos A., Magkou C., Bakarakos P., Nakopoulou L. The clinicopathologic and prognostic significance of CD44+/CD24(-/low) and CD44-/CD24+ tumor cells in invasive breast carcinomas. Hum Pathol. 2008 Jul; 39(7): 1096–102. doi: 10.1016/j.humpath.2007.12.003.

16. Lü X., Xu K., Lü H., Yin Y., Ma C., Liu Y., Li H., Suo Z. CD44(+)/ CD24(-) cells are transit progenitors and do not determine the molecular subtypes and clinical parameters in breast carcinomas. Ultrastruct Pathol. 2011 Apr; 35(2): 72–8. doi: 10.3109/01913123.2010.544843.

17. Uchôa Dde M., Graudenz M.S., Callegari-Jacques S.M., Hartmann C.R., Ferreira B.P., Fitarelli-Kiehl M., Edelweiss M.I. Expression of cancer stem cell markers in basal and penta-negative breast carcinomas--a study of a series of triple-negative tumors. Pathol Res Pract. 2014 Jul; 210(7): 432–9. doi: 10.1016/j.prp.2014.03.005.

18. Yang F., Cao L., Sun Z., Jin J., Fang H., Zhang W., Guan X. Evaluation of Breast Cancer Stem Cells and Intratumor Stemness Heterogeneity in Triple-negative Breast Cancer as Prognostic Factors. Int J Biol Sci. 2016 Dec 7; 12(12): 1568–1577. doi: 10.7150/ijbs.16874.

19. Matchuk O.N., Zamulaeva I.A., Kovalev O.A., Saenko A.S. Radioresistance mechanisms of side population cells in mouse melanoma cell line B16. Tsitologiia. 2013; 55(8): 553–9. (in Russian).

20. Chen Y., Song J., Jiang Y., Yu C., Ma Z. Predictive value of CD44 and CD24 for prognosis and chemotherapy response in invasive breast ductal carcinoma. Int J Clin Exp Pathol. 2015; 8(9): 11287–11295.

21. Kapucuoğlu N., Bozkurt K.K., Başpınar Ş., Koçer M., Eroğlu H.E., Akdeniz R., Akçil M. The clinicopathological and prognostic significance of CD24, CD44, CD133, ALDH1 expressions in invasive ductal carcinoma of the breast: CD44/CD24 expression in breast cancer. Pathol Res Pract. 2015 Oct; 211(10): 740–7. doi: 10.1016/j.prp.2015.05.011.

22. Collina F., Di Bonito M., Li Bergolis V., De Laurentiis M., Vitagliano C., Cerrone M., Nuzzo F., Cantile M., Botti G. Prognostic Value of Cancer Stem Cells Markers in Triple-Negative Breast Cancer. Biomed Res Int. 2015; 2015: 158682. doi: 10.1155/2015/158682.