Abstract
Breast cancer is the most common malignancy worldwide in females, representing 29% of all cancer new cases and 14% of cancer deaths in the world. Amongst the reasons for the high mortality rate is resistance to chemotherapy resulting in therapeutic failure. Various studies have shown that the presence of cancer stem cells (CSCs) in breast tumors is responsible for chemotherapy resistance and tumor recurrence. This CSC population possesses the characteristics of normal stem cells, including their ability to self-renewal and give rise to other epithelial cells. One thing that unique to the CSC population is their ability to escape from chemotherapy drugs; this can make them resistant to therapy and able to repopulate the cancer. Isolation and enrichment of breast CSCs (BCSCs) is required in order to study their characteristics and the behavior that enables them to drive breast tumor development, in order to develop better therapies. This chapter describes a method for the isolation and enrichment of BCSCs from the MCF7 breast cancer cell line, which consists of a heterogeneous breast cancer cell population. This method depends on cancer stem cell behavior, specifically an ability to self-renew and form spheroids in harsh conditions that allow only cancer cells with stem cell characteristics to survive and form spheroids.
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References
Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A 100(7):3983–3988
Ghanei Z, Jamshidizad A, Joupari MD, Shamsara M (2020) Isolation and characterization of breast cancer stem cell-like phenotype by Oct4 promoter-mediumted activity. J Cell Physiol. [cited 2020 Apr 8]. http://www.ncbi.nlm.nih.gov/pubmed/31904128
Wang H, Zhang Y, Du Y (2013) Ovarian and breast cancer spheres are similar in transcriptomic features and sensitive to fenretinide. Biomed Res Int 2013:510905
Saadin K, White IM (2013) Breast cancer stem cell enrichment and isolation by mammosphere culture and its potential diagnostic applications. Expert Rev Mol Diagn 13:49–60
Harrison H, Shaw FL, Harrison H, Spence K, Ablett MP, Simões BM et al (2012) A detailed mammosphere assay protocol for the quantification of breast stem cell activity. J Mammary Gland Biol Neoplasia 17(2):111–117
Sin WC, Lim CL (2017) Breast cancer stem cells—from origins to targeted therapy. Stem Cell Investig 6
Velasco-Velázquez MA, Popov VM, Lisanti MP, Pestell RG (2011) The role of breast cancer stem cells in metastasis and therapeutic implications. Am J Pathol Am Soc Investig Pathol 179:2–11
Geng SQ, Alexandrou AT, Li JJ (2014) Breast cancer stem cells: multiple capacities in tumor metastasis. Cancer Lett 349:1–7. [cited 2020 Apr 8]. http://www.ncbi.nlm.nih.gov/pubmed/24727284
Fico F, Bousquenaud M, Rüegg C, Santamaria-MartÃnez A (2019) Breast cancer stem cells with tumor- versus metastasis-initiating capacities are modulated by TGFBR1 inhibition. Stem Cell Rep 13(1):1–9
Ayob AZ, Ramasamy TS (2018) Cancer stem cells as key drivers of tumour progression. J Biomed Sci 25
Park SY, Choi JH, Nam JS (2019) Targeting cancer stem cells in triple-negative breast cancer. Cancers 11
Pastrana E (2013) Eyes wide open: a critical review of sphere-formation as an assay for stem cells. Cell Stem Cell 8(5):486–498
Bao B, Ahmad A, Azmi AS, Ali S, Sarkar FH (2013) Overview of cancer stem cells (CSCs) and mechanisms of their regulation: implications for cancer therapy. Curr Protoc Pharmacol 61(1):14.25.1–14.25.14. [cited 2020 May 2]
Zhang J, Zhang Y, Cheng L, Li C, Dai L, Zhang H et al (2018) Enrichment and characterization of cancer stem-like cells in ultra-low concentration of serum and non-adhesive culture system. Am J Transl Res 10(5):1552–1561
Lee CH, Yu CC, Wang BY, Chang WW (2016) Tumorsphere as an effective in vitro platform for screening anticancer stem cell drugs. Oncotarget 7(2):1215–1226
Herheliuk T, Perepelytsina O, Ugnivenko A, Ostapchenko L, Sydorenko M (2019) Investigation of multicellular tumor spheroids enriched for a cancer stem cell phenotype. Stem Cell Investig 6
Ishiguro T, Ohata H, Sato A, Yamawaki K, Enomoto T, Okamoto K (2017) Tumor-derived spheroids: relevance to cancer stem cells and clinical applications. Cancer Sci 108:283–289
Zhong Y, Guan K, Guo S, Zhou C, Wang D, Ma W et al (2010) Spheres derived from the human SK-RC-42 renal cell carcinoma cell line are enriched in cancer stem cells. Cancer Lett 299(2):150–160
Lee JW (2018) Isolation of spheroid-forming single cells from gastric cancer cell lines: enrichment of cancer stem-like cells. BioTechniques 65(4):197–203
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Ishtiah, A.A., Yahaya, B.H. (2022). The Enrichment of Breast Cancer Stem Cells from MCF7 Breast Cancer Cell Line Using Spheroid Culture Technique. In: Kannan, N., Beer, P. (eds) Stem Cell Assays. Methods in Molecular Biology, vol 2429. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1979-7_32
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DOI: https://doi.org/10.1007/978-1-0716-1979-7_32
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Publisher Name: Humana, New York, NY
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