Selection for EGFR gene amplification in a breast epithelial cell line with basal-like phenotype and hereditary background

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. Immun...

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Bibliographic Details
Published in:In vitro cellular & developmental biology. Animal 2011-02, Vol.47 (2), p.139-148
Main Authors: Ingthorsson, Saevar, Halldorsson, Thorhallur, Sigurdsson, Valgardur, Friðriksdottir, Agla JR, Bodvarsdottir, Sigridur K., Steinarsdottir, Margret, Johannsson, Oskar, Magnusson, Magnus K., Ogmundsdottir, Helga M., Gudjonsson, Thorarinn
Format: Article
Language:English
Subjects:
Animal Genetics and Genomics
Antibodies
Basement membranes
Biomedical and Life Sciences
Breast cancer
Breast carcinoma
Breast Neoplasms - genetics
Breast Neoplasms - pathology
Carcinoma - genetics
Carcinoma - pathology
CELL AND TISSUE MODELS
Cell Biology
Cell Culture
Cell culture techniques
Cell growth
Cell Line, Tumor
Cell lines
Cell Proliferation
Chromosomes
Contact inhibition
Culture Media, Serum-Free - pharmacology
Cultured cells
Cytokeratin
Developmental Biology
Epidermal growth factor
Epidermal Growth Factor - pharmacology
Epidermal growth factor receptors
Epithelial cells
Epithelial Cells - drug effects
Epithelial Cells - pathology
Female
Filters
Fluorescence in situ hybridization
Gene Amplification
Humans
Karyotypes
Life Sciences
Middle Aged
Mutation
Neoplasms, Basal Cell - genetics
Neoplasms, Basal Cell - pathology
Phenotypes
Polarity
Prognosis
Receptor, Epidermal Growth Factor - genetics
Selection, Genetic
Stem Cells
Tumors
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Summary: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 sub-population 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.
ISSN:1071-2690
1543-706X
DOI:10.1007/s11626-010-9371-6