Molecular and Cellular Biochemistry

, Volume 306, Issue 1–2, pp 201–212 | Cite as

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

  • Matthias Christgen
  • Matthias Ballmaier
  • Henriette Bruchhardt
  • Reinhard von Wasielewski
  • Hans Kreipe
  • Ulrich Lehmann
Article

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.

Keywords

Cancer stem cells Differentiation Intratumoral heterogeneity BCRP1 Cal51 

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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Matthias Christgen
    • 1
  • Matthias Ballmaier
    • 2
  • Henriette Bruchhardt
    • 1
  • Reinhard von Wasielewski
    • 1
  • Hans Kreipe
    • 1
  • Ulrich Lehmann
    • 1
  1. 1.Institute of PathologyHannover Medical SchoolHannoverGermany
  2. 2.Department of Pediatric HaematologyHannover Medical SchoolHannoverGermany

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