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. 2020 Nov;235(11):7840-7848.
doi: 10.1002/jcp.29437. Epub 2020 Jan 6.

Isolation and characterization of breast cancer stem cell-like phenotype by Oct4 promoter-mediated activity

Zahra Ghanei  1 , Abbas Jamshidizad  1 , Morteza Daliri Joupari  1 , Mehdi Shamsara  1
Affiliations

Isolation and characterization of breast cancer stem cell-like phenotype by Oct4 promoter-mediated activity

Zahra Ghanei et al. J Cell Physiol. 2020 Nov.

Abstract

Cancer stem cells (CSCs) are a small subset of cancer cells responsible for self-renewal activity, drug resistance, and tumor recurrence. CSCs have been derived from diverse tumors and cell lines. The expression of stemness markers has been identified in CSCs. Oct4 is a well-established transcription factor expressed in stem cells and CSCs. In this study, we isolated and characterized breast CSC-like cells from murine MC4-L2 cells by Oct4 promoter-mediated activity. The MC4-L2 cells were electroporated by a plasmid expressing puromycin resistance (PuroR ) gene from the Oct4 promoter and then selected by puromycin. The isolated cells were named as the MC4-L2puro cells and characterized for CSCs properties. Immunostaining indicated CD44high and CD24high phenotype for the MC4-L2 and MC4-L2puro cells. The enhanced expression of stem cell markers was detected in the puromycin-selected cells compared with the parental cells. Moreover, the isolated cells only grew up in sphere-formed shape in low attachment plates. Serial dilution transplantation in syngeneic mouse models showed increased tumorigenicity of the MC4-L2puro cells, as they induced new tumors when injected into the mammary fat pad as few as 104 cells. In conclusion, we designed a novel genetic construct, which allows the isolation of Oct4-positive cells in a cancer population by a simple selection step in a puromycin-containing medium. Transfection of this construct into the MC4-L2 cells resulted in growing a subpopulation of cells having tumor-initiating cell characteristics. To the best of our knowledge, this is the first report on the isolation of CSC-like cells from the mouse breast cancer MC4-L2 cells.

Keywords: MC4-L2 cells; Oct4; breast cancer; cancer stem cell; mouse.

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References

REFERENCES

    1. Abbaszadegan, M. R., Bagheri, V., Razavi, M. S., Momtazi, A. A., Sahebkar, A., & Gholamin, M. (2017). Isolation, identification, and characterization of cancer stem cells: A review. Journal of Cellular Physiology, 232(8), 2008-2018. https://doi.org/10.1002/jcp.25759
    1. Abubaker, K., Latifi, A., Luwor, R., Nazaretian, S., Zhu, H., Quinn, M. A., … Ahmed, N. (2013). Short-term single treatment of chemotherapy results in the enrichment of ovarian cancer stem cell-like cells leading to an increased tumor burden. Molecular Cancer, 12(1), 24. https://doi.org/10.1186/1476-4598-12-24
    1. Ajani, J. A., Song, S., Hochster, H. S., & Steinberg, I. B. (2015). Cancer stem cells: The promise and the potential. Seminars in Oncology, 42(Suppl. 1), S3-S17. https://doi.org/10.1053/j.seminoncol.2015.01.001
    1. Al-Hajj, M., Wicha, M. S., Benito-Hernandez, A., Morrison, S. J., & Clarke, M. F. (2003). Prospective identification of tumorigenic breast cancer cells. Proceedings of the National Academy of Sciences of the United States of America, 100(7), 3983-3988. https://doi.org/10.1073/pnas.0530291100
    1. Batlle, E., & Clevers, H. (2017). Cancer stem cells revisited. Nature Medicine, 23(10), 1124-1134. https://doi.org/10.1038/nm.4409

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