SENSITIVITY OF BREAST CANCER STEM CELLS (CD24-/CD44+) TO DOXORUBICIN IS ASSOCIATED WITH OXIDATIVE STRESS STATUS
Objective: The aim of this study was to analyze the sensitivity of BCSCs to doxorubicin and its association with oxidative stress.
Methods: BCSCs (CD24-/CD44+) were treated with doxorubicin every 2 d for 14 d. The determination of cell viability was performed using a trypan blue exclusion assay. The levels of reactive oxygen species (ROS) were measured using a dihydroethidium (DHE) and a 2’,7’-dichlorofluorescein diacetate (DCFH-DA) probes. Manganese superoxide dismutase (MnSOD) mRNA expression and specific activity were also analyzed. Glutathione (GSH) level was measured using Ellman’s method.
Results: The viability of the BCSCs decreased after 2 d of treatment with doxorubicin, but started to increase after 8 d. After 8 d of doxorubicin treatment, the ROS level in the BCSCs decreased, while the MnSOD specific activity increased. In addition, the MnSOD mRNA expression and GSH level were suppressed after 8 d of treatment.
Conclusion: Doxorubicin treatment induced cytotoxicity after 2 d by increasing the superoxide levels of the BCSCs. After 8 d of treatment, the sensitivity of BCSCs to doxorubicin decreased due to the suppressed oxidative stress from the enhanced antioxidant activity of the MnSOD.
2. Nounou MI, ElAmrawy F, Ahmed N, Abdelraouf K, Goda S, Syed-Sha-Qhattal H. Breast cancer: conventional diagnosis and treatment modalities and recent patents and technologies. Breast Cancer: Basic Clin Res 2015;9:17-34.
3. Ahmad A. Pathways to breast cancer recurrence. ISRN Oncology; 2013. p. 1-16.
4. Zheng S, Xin L, Liang A, Fu Y. Cancer stem cell hypothesis: a brief summary and two proposals. Cytotechnology 2013;65:505-12.
5. Nassar D, Blanpain C. Cancer stem cells: basic concepts and therapeutic implications. Annu Rev Pathol: Mech Dis 2016;11:47-76.
6. Takahashi R, Takeshita F, Fujiwara T, Ono M, Ochiya T. Cancer stem cells in breast cancer. Cancers 2011;3:1311-28.
7. Li W, Ma H, Zhang J, Zhu L, Wang C, Yang Y. Unraveling the roles of CD44/CD24 and ALDH1 as cancer stem cell markers in tumorigenesis and metastasis. Sci Rep 2017;7:1-15.
8. Horimoto Y, Arakawa A, Sasahara N, Tanabe M, Sai S, Himuro T, et al. Combination of cancer stem cell markers CD44 and CD24 is superior to ALDH1 as a prognostic indicator in breast cancer patients with distant metastases. PLoS ONE 2016;11:1-11.
9. Al-Ejeh F, Smart CE, Morrison BJ, Chenevix Trench G, Lopez JA, Lakhani SR, et al. Breast cancer stem cells: treatment resistance and therapeutic opportunities. Carcinogenesis 2011;32:650-8.
10. Lovitt CJ, Shelper TB, Avery VM. Doxorubicin resistance in breast cancer cells is mediated by extracellular matrix proteins. BMC Cancer 2018;18:41.
11. Carvalho C, Santos RX, Cardoso S, Correia S, Oliveira PJ, Santos MS, et al. Doxorubicin: the good, the bad, and the ugly effect. Curr Med Chem 2009;16:3267-85.
12. Thorn CF, Oshiro C, Marsh S, Hernandez Boussard T, McLeod H, Klein TE, et al. Doxorubicin pathways: pharmacodynamics and adverse effects. Pharmacogenet Genomics 2011;21:440-6.
13. Abu Hammad S, Zihlif M. Gene expression alterations in doxorubicin resistant MCF7 breast cancer cell line. Genomics 2013;101:213-20.
14. Mencalha A, Victorino VJ, Cecchini R, Panis C. Mapping oxidative changes in breast cancer: understanding the basic to reach the clinics. Anticancer Res 2014;34:1127-40.
15. Fu A, Ma S, Wei N, Tan BXX, Tan EY, Luo KQ. High expression of MnSOD promotes survival of circulating breast cancer cells and increases their resistance to doxorubicin. Oncotarget 2016;7:50239-57.
16. Jardim BV, Moschetta MG, Leonel C, Gelaleti GB, Regiani VR, Ferreira LC, et al. Glutathione and glutathione peroxidase expression in breast cancer: an immunohistochemical and molecular study. Oncol Reports 2013;30:1119-28.
17. Traverso N, Ricciarelli R, Nitti M, Marengo B, Furfaro A, Prontazo M, et al. Role of glutathione in cancer progression and chemoresistance. Oxidative Med Cellular Longevity 2013;1-10. http://dx.doi.org/10.1155/2013/972913
18. Wanandi SI, Yustisia I, Neolaka GMG, Jusman SWA. Impact of extracellular alkalinization on the survival of human CD24-/CD44+breast cancer stem cells associated with cellular metabolic shifts. Brazilian J Med Biological Res 2017;50:1-11.
19. Wanandi SI, Syahrani RA, Jusman SWA. The impact of rotenone-modulated oxidative stress on the survival of human breast cancer stem cells (CD24-/CD44+). Ann Oncol 2017;28. https://doi.org/10.1093/annonc/mdx361.007
20. Hardiany NS, Sadikin M, Siregar N, Wanandi SI. The suppression of manganese superoxide dismutase decreased the survival of human glioblastoma multiforme T98G cells. Med J Indonesia 2017;26:19-25.
21. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 2001:25:402-8.
22. Laragione T, Bonetto V, Casoni F, Massignan T, Bianchi G, Gianazza E, et al. Redox regulation of surface protein thiols: identification of integrin ?-4 as a molecular target by using redox proteomics. Proc Natl Acad Sci USA 2003;100:14737-41.
23. Coley MH. Mechanisms and consequences of chemotherapy resistance in breast cancer. Eur J Cancer Suppl 2009;7:3-7.
24. Louisa M, Soediro TM, Suyatna FD. In vitro modulation of P-glycoprotein, MRP-1 and BCRP expression by mangiferin in doxorubicin-treated MCF-7 cells. Asian Pac J Cancer Prev 2014;14:1639-42.
25. Yenigun VB, Ozpolat B, Kose GT. Response of CD44+/CD24-/low breast cancer stem/progenitor cells to tamoxifen-and doxorubicin-induced autophagy. Int J Mol Med 2013;31:1477-83.
26. Shi X, Zhang Y, Zheng J, Pan J. Reactive oxygen species in cancer stem cells. Antioxid Redox Signaling 2012;16:1215-28.
27. Diehn M, Cho RW, Lobo NA, Kalisky T, Dorie MJ, et al. Association of reactive oxygen species levels and radioresistance in cancer stem cells. Nature 2009;458:780-3.
28. Wei ZB, Miao XY, Yang MQ, Luo XG. Advances in the espression and regulation of MnSOD gene. Yi Chuan 2008;30:831-7.
29. Becuwe P, Ennen M, Klotz R, Barbieux C, Grandemange S. Manganese superoxide dismutase in breast cancer: from molecular mechanisms of gene regulation to biological and clinical significance. Free Radical Biol Med 2014;77:139-51.
30. Candas D, Li JJ. MnSOD in oxidative stress response-potential regulation via mitochondrial protein influx. Antioxid Redox Signaling 2014;20:1599-617.
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