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Identification of a distinct side population of cancer cells in the Cal-51 human breast carcinoma cell line

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Abstract

“Side population” (SP) cells, which pump out the fluorescent dye H33342 via the ABCG2 transporter, define a putative stem/progenitor cell population in the mammary gland. Breast cancer SP cells recently isolated from the MCF-7 cell line possess similar properties and may represent stem cell-like cancer cells. This study extends SP cell analysis to a broad panel of human breast cancer cell lines and investigates the expression of differentiation-associated markers in isolated cancer SP cells. Expression of ABCG2 was determined in 16 breast cancer cell lines by quantitative RT-PCR, Western blotting and immunohistochemistry. Subsequently, all cell lines were screened for the presence of SP cells. Human breast cancer cell lines commonly express ABCG2. ABCG2-immunoreactivity was clearly restricted to rare cancer cells in several cell lines including Cal-51. Analysis of H33342-labeled Cal-51 cells revealed a small fraction of putative SP cells accounting for one percent of all cells. The genuine nature of Cal-51 SP cells was unambiguously verified by demonstrating a 30-fold increased ABCG2-expression in isolated Cal-51 SP cells. During in vitro expansion, Cal-51 SP cells generated heterologous non-SP (NSP) cells and ABCG2-expression declined dramatically. In contrast, NSP cells failed to sustain proliferation. Freshly isolated Cal-51 SP cells also exhibited increased expression of Muc1 and CALLA. Noteworthy, non-malignant mammary epithelial SP cells lack these differentiation markers, highlighting fundamental differences between non-malignant and breast cancer-derived SP cells. In summary, we established Cal-51 SP cells as a novel in vitro model to study differential gene expression in breast cancer-derived SP and NSP cells.

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Authors and Affiliations

  1. Institute of Pathology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Matthias Christgen, Henriette Bruchhardt, Reinhard von Wasielewski, Hans Kreipe & Ulrich Lehmann

  2. Department of Pediatric Haematology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Matthias Ballmaier

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  1. Matthias Christgen
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  2. Matthias Ballmaier
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  3. Henriette Bruchhardt
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  4. Reinhard von Wasielewski
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  5. Hans Kreipe
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  6. Ulrich Lehmann
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Correspondence to Matthias Christgen.

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Christgen, M., Ballmaier, M., Bruchhardt, H. et al. Identification of a distinct side population of cancer cells in the Cal-51 human breast carcinoma cell line. Mol Cell Biochem 306, 201–212 (2007). https://doi.org/10.1007/s11010-007-9570-y

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  • DOI: https://doi.org/10.1007/s11010-007-9570-y

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