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Establishment, growth and in vivo differentiation of a new clonal human cell line, EM-G3, derived from breast cancer progenitors

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Abstract

A new clonal cell line, EM-G3, was derived from a primary lesion of human infiltrating ductal breast carcinoma. The line consisted of cuboidal cells with occasional appearance of more differentiated branched cells apparently involved in cell-to-cell communication. The EM-G3 cells, population doubling time 34 h, are dependent on the epidermal growth factor. Multicolor fluorescence in situ hybridization (mFISH) analysis demonstrated a stable diploid genome with several genetic changes. Immunocytochemical analysis of EM-G3 in vitro revealed positivity for keratins (K) K5, K14, K18, nuclear protein p63, epithelial membrane antigen (EMA) and other proteins indicative of a pattern of mammary epithelium bipotent progenitors. Detection of integrins α-6, β-1, and protein CD44 by cDNA array also pointed to the character of basal/stem cells. In contrast, dominant cells in the human original tumor showed the luminal character (K18+, K19+, K5−, K14−, and p63−). However, cells with the immunocytochemical profile similar to that of cultured EM-G3 cells were found in minor clusters in the patient’s tumor sections. The EM-G3 cells formed limited tumors in nu/nu mice. The cells in mouse tumors were organized in primitive ductal-like structures consisting of 1–3 large central luminal-like cells (EMA+) surrounded by peripheral myoepithelial-like cells (p63+/EMA−). The large central cells gradually disintegrated, forming a pseudolumen. Apparently, EM-G3 cells are able to partially differentiate in vivo as well as in vitro. Our results indicate that EM-G3 cells were derived from a premalignant population of common progenitors of luminal and myoepithelial cells that were immortalized in an early stage of tumorigenesis.

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Acknowledgments

We thank Mrs. Eva Taislova for excellent technical assistance. This work was supported by grants NR8145–3 and NR8345–4 from the Grant Agency of the Ministry of Health of the Czech Republic, Grant No. 93/2005/C from the Grant Agency of Charles University, Project AVOZ50520514 from the Academy of Sciences of the Czech Republic, and Research Project MSM0021620808 from the Ministry of Education, Youth and Sports of the Czech Republic

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Authors and Affiliations

  1. Department of Cell Biology, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nam. 2, 166 37, Prague 6, Czech Republic

    Marketa Brozova, Alena Chaloupkova, Pavel Vesely, Luboslava Krasna & Eva Matouskova

  2. First Faculty of Medicine, Institute of Biochemistry and Experimental Oncology, Charles University in Prague, Prague, Czech Republic

    Zdenek Kleibl, Jan Sevcik & Eva Scholzova

  3. General Faculty Hospital in Prague, Laboratory of Cytostatics, Prague, Czech Republic

    Irena Netikova

  4. Institute of Hematology and Blood Transfusion, Prague, Czech Republic

    Jana Brezinova

  5. First Faculty of Medicine, Department of Pathology, Charles University in Prague, Prague, Czech Republic

    Pavel Dundr

  6. First Faculty of Medicine, Institute of Biophysics and Informatics, Charles University in Prague, Prague, Czech Republic

    Marie Zadinova

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  1. Marketa Brozova
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Correspondence to Eva Matouskova.

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Brozova, M., Kleibl, Z., Netikova, I. et al. Establishment, growth and in vivo differentiation of a new clonal human cell line, EM-G3, derived from breast cancer progenitors. Breast Cancer Res Treat 103, 247–257 (2007). https://doi.org/10.1007/s10549-006-9358-x

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  • DOI: https://doi.org/10.1007/s10549-006-9358-x

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