Abstract
An epithelial cell line, referred to as A163, was established from breast carcinoma derived from a patient with a strong family history of breast cancer but no known breast cancer susceptibility mutation. A163 was propagated in a serum-free culture medium including the epidermal growth factor. Immunophenotypic characterization demonstrated a mixed luminal and basal-like phenotype. When epidermal growth factor was excluded from the culture medium, A163 entered a quiescent period followed by a period of increased cell proliferation in a subpopulation of the cells. The epidermal growth factor-independent subpopulation retained the basal-like phenotype of the parental cell line. Karyotype and fluorescent in situ hybridization analysis showed an amplification of epidermal growth factor receptor on 7q in A163-S1 only, resulting in high expression of total and phosphorylated epidermal growth factor receptor. The A163-S1 sub-line piles up in culture, indicating a loss of contact inhibition. When grown on transwell filters, A163 shows basal expression of P63 and cytokeratin 14, whereas A163-S1 expresses P63 ubiquitously, and has lost the basal specific expression of cytokeratin 14, indicating a loss of polarity. Furthermore, when cultured in reconstituted basement membrane matrix, A163 form polarized normal like acini. In contrast, A163-S1 form large disorganized structures with lack of polarity. These cell lines may prove useful to understand molecular changes in breast cancer progression, in particular basal-like breast cancer subtype with bad prognosis and no current treatment options.
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Acknowledgments
Grant support was provided by the Icelandic Research Council, Landspitali University Hospital Research Fund, University of Iceland Research Fund, Science and Technology Policy Council-Thematic program in postgenomic biomedicine. European Science Foundation (EuroCORES program, EuroSTELLS), “Göngum saman” a supporting group for breast cancer research in Iceland. This work has been approved by the National Bioethics Committee of Iceland, Reference number VSNa2001050056.
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Editor: J. Denry Sato
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Ingthorsson, S., Halldorsson, T., Sigurdsson, V. et al. Selection for EGFR gene amplification in a breast epithelial cell line with basal-like phenotype and hereditary background. In Vitro Cell.Dev.Biol.-Animal 47, 139–148 (2011). https://doi.org/10.1007/s11626-010-9371-6
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DOI: https://doi.org/10.1007/s11626-010-9371-6