Irradiation and conditioned media from human umbilical cord stem cells suppress epithelial-mesenchymal transition biomarkers in breast cancer cells

Document Type : Original Article


1 Department of Medical Physics, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

2 Cellular and Molecular Research Center, Medical Basic Sciencec Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

3 Department of Anatomical Sciences, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

4 Department of Virology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran



Objective(s): Breast cancer cells developing radioresistance during radiation may result in cancer recurrence and poor survival. One of the main reasons for this problem is the changes in the regulation of genes that have a key role in the epithelial-mesenchymal transition (EMT). Utilizing mesenchymal stem cells can be an effective approach to overcome therapeutic resistance. In this study, we investigated the possibility of combining mesenchymal medium with cancer cell medium in sensitizing breast carcinoma cells to radiation. 
Materials and Methods: In this experimental study, the cells were irradiated at a dose of 4 Gy alone and in combination with stem cells and cancer cells media. Apoptosis, cell cycle, Western blotting, and real-time PCR assays evaluated the therapeutic effects.  
Results: We found that the CSCM could decrease the expression of several EMT markers (CD133, CD44, Vimentin, Nanog, Snail, and Twist), resulting in increased cell distribution in the G1 and G2/M phases, apoptosis rate, and protein levels of p-Chk2 and cyclin D1; furthermore, it exhibits synergetic effects with radiation treatment in vitro. 
Conclusion: These findings show that CSCM inhibits the expansion of breast cancer cells and makes them more susceptible to radiotherapy, offering a unique approach to treating breast cancer by overcoming radioresistance.


1. Chavez KJ, Garimella SV, Lipkowitz S. Triple negative breast cancer cell lines: One tool in the search for better treatment of triple negative breast cancer. Breast Dis 2010; 32:35-48.
2. Hashemitabar M, Allahbakhshi E, Tabande MR, Orazizadeh M, Dehbashi FN, Azandeh S, et al. Isolation and characterization of human umbilical cord mesenchymal stem cells and their differentiation into Pdx-1+ Cells. J Biomed Sci Eng 2015; 8:780.
3. Gudkov AV, Komarova EA. The role of p53 in determining sensitivity to radiotherapy. Nat Rev Cancer 2003; 3:117-129.
4. Jia-Quan Q, Hong-Mei Y, Xu Y, Li-Na L, Jin-Feng Z, Ta X, et al. MiR-23a sensitizes nasopharyngeal carcinoma to irradiation by targeting IL-8/Stat3 pathway. Oncotarget 2015; 6:28341.
5. Brown JM, Wouters BG. Apoptosis, p53, and tumor cell sensitivity to anticancer agents. Cancer Res 1999; 59:1391-1399.
6. Wang X, Lin Y. Tumor necrosis factor and cancer, buddies or foes? Acta Pharmacol Sin 2008; 29:1275-1288.
7. Zhang Z, Lin G, Yan Y, Li X, Hu Y, Wang J, et al. Transmembrane TNF-alpha promotes chemoresistance in breast cancer cells. Oncogene 2018; 37:3456-3470.
8. Yu M, Zhou X, Niu L, Lin G, Huang J, Zhou W, et al. Targeting transmembrane TNF-alpha suppresses breast cancer growth. Cancer Res 2013; 73:4061-4074.
9. Chakraborty S, Rahman T. The difficulties in cancer treatment. Ecancermedicalscience 2012; 6:ed16.
10. De Bacco F, Luraghi P, Medico E, Reato G, Girolami F, Perera T, et al. Induction of MET by ionizing radiation and its role in radioresistance and invasive growth of cancer. J Natl Cancer Inst 2011; 103:645-661.
11. Wild-Bode C, Weller M, Rimner A, Dichgans J, Wick W. Sublethal irradiation promotes migration and invasiveness of glioma cells: implications for radiotherapy of human glioblastoma. Cancer Res 2001; 61:2744-2750.
12. Ramasamy R, Lam EW, Soeiro I, Tisato V, Bonnet D, Dazzi F. Mesenchymal stem cells inhibit proliferation and apoptosis of tumor cells: Impact on in vivo tumor growth. Leukemia 2007; 21:304-310.
13. Lin HD, Fong CY, Biswas A, Choolani M, Bongso A. Human umbilical cord wharton’s jelly stem cell conditioned medium induces tumoricidal effects on lymphoma cells through hydrogen peroxide mediation. J Cell Biochem 2016; 117:2045-2055.
14. Brabletz T, Kalluri R, Nieto MA, Weinberg RA. EMT in cancer. Nat Rev Cancer 2018; 18:128-134.
15. Lamouille S, Xu J, Derynck R. Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol 2014; 15:178-196.
16. Vasefifar P, Motafakkerazad R, Maleki LA, Najafi S, Ghrobaninezhad F, Najafzadeh B, et al. Nanog, as a key cancer stem cell marker in tumor progression. Gene 2022; 827:146448.
17. Weinberg RA, Weinberg RA. The Biology of Cancer. 1st ed: WW Norton & Company; 2006.
18. Hazan RB, Phillips GR, Qiao RF, Norton L, Aaronson SA. Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis. J Cell Biol 2000; 148:779-790.
19. Kim R-K, Kaushik N, Suh Y, Yoo K-C, Cui Y-H, Kim M-J, et al. Radiation driven epithelial-mesenchymal transition is mediated by Notch signaling in breast cancer. Oncotarget 2016; 7:53430.
20. Ulirsch J, Fan C, Knafl G, Wu MJ, Coleman B, Perou CM, et al. Vimentin DNA methylation predicts survival in breast cancer. Breast Cancer Res Treat 2013; 137:383-396.
21. Serej ZA, Ebrahimi A, Kazemi T, Najafi S, Amini M, Nastarin P, et al. NANOG gene suppression and replacement of let-7 modulate the stemness, invasion, and apoptosis in breast cancer. Gene 2021; 801:145844.
22. Zhu QQ, Ma C, Wang Q, Song Y, Lv T. The role of TWIST1 in epithelial-mesenchymal transition and cancers. Tumour Biol 2016; 37:185-197.
23. Vesuna F, Bergman Y, Raman V. Genomic pathways modulated by Twist in breast cancer. BMC cancer 2017; 17:1-12.
24. Kurrey NK, Jalgaonkar SP, Joglekar AV, Ghanate AD, Chaskar PD, Doiphode RY, et al. Snail and slug mediate radioresistance and chemoresistance by antagonizing p53-mediated apoptosis and acquiring a stem-like phenotype in ovarian cancer cells. Stem Cells 2009; 27:2059-2068.
25. Lin Y, Bai X, Zhou W, He Y, Wu Y, Wang X. Radiation exposure triggers the progression of triple negative breast cancer via stabilizing ZEB1. Biomed Pharmacother 2018; 107:1624-1630.
26. Chiou SH, Kao CL, Chen YW, Chien CS, Hung SC, Lo JF, et al. Identification of CD133-positive radioresistant cells in atypical teratoid/rhabdoid tumor. PLoS One 2008; 3:e2090.
27. Ke CC, Liu RS, Yang AH, Liu CS, Chi CW, Tseng LM, et al. CD133-expressing thyroid cancer cells are undifferentiated, radioresistant and survive radioiodide therapy. Eur J Nucl Med Mol Imaging 2013; 40:61-71.
28. He Y, Liu Z, Qiao C, Xu M, Yu J, Li G. Expression and significance of Wnt signaling components and their target genes in breast carcinoma. Mol Med Rep 2014; 9:137-143.
29. Arnold A, Papanikolaou A. Cyclin D1 in breast cancer pathogenesis. J Clin Oncol 2005; 23:4215-4224.
30. Yin H, Glass J. The phenotypic radiation resistance of CD44+/CD24- or low breast cancer cells is mediated through the enhanced activation of ATM signaling. PLoS One 2011; 6:e24080.