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01.10.2010 | Research article | Ausgabe 5/2010 Open Access

Breast Cancer Research 5/2010

Mapping the cellular and molecular heterogeneity of normal and malignant breast tissues and cultured cell lines

Zeitschrift:
Breast Cancer Research > Ausgabe 5/2010
Autoren:
Patricia J Keller, Amy F Lin, Lisa M Arendt, Ina Klebba, Ainsley D Jones, Jenny A Rudnick, Theresa A DiMeo, Hannah Gilmore, Douglas M Jefferson, Roger A Graham, Stephen P Naber, Stuart Schnitt, Charlotte Kuperwasser
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​bcr2755) contains supplementary material, which is available to authorized users.
Patricia J Keller, Amy F Lin contributed equally to this work.

Competing interests

The authors declare that they have no competing interests.

Authors' contributions

PJK, AL, LMA and ADJ took part in the conception and design of the study, the collection of data, data analysis and interpretation, and manuscript writing. IK collected and/or assembled data. CF and CMP took part in the collection of data, and data analysis and interpretation. JAR and TAD collected data. HG, SS, RAG, DJ and SN dealt with the provision of study materials, including the procurement of resources and samples (cell lines or reduction mammoplasty and tumor tissues). CK took part in the conception and design of the study, the collection and/or assembly of data, data analysis and interpretation, manuscript writing and financial support. All authors approved of the final manuscript.

Abstract

Introduction

Normal and neoplastic breast tissues are comprised of heterogeneous populations of epithelial cells exhibiting various degrees of maturation and differentiation. While cultured cell lines have been derived from both normal and malignant tissues, it remains unclear to what extent they retain similar levels of differentiation and heterogeneity as that found within breast tissues.

Methods

We used 12 reduction mammoplasty tissues, 15 primary breast cancer tissues, and 20 human breast epithelial cell lines (16 cancer lines, 4 normal lines) to perform flow cytometry for CD44, CD24, epithelial cell adhesion molecule (EpCAM), and CD49f expression, as well as immunohistochemistry, and in vivo tumor xenograft formation studies to extensively analyze the molecular and cellular characteristics of breast epithelial cell lineages.

Results

Human breast tissues contain four distinguishable epithelial differentiation states (two luminal phenotypes and two basal phenotypes) that differ on the basis of CD24, EpCAM and CD49f expression. Primary human breast cancer tissues also contain these four cellular states, but in altered proportions compared to normal tissues. In contrast, cultured cancer cell lines are enriched for rare basal and mesenchymal epithelial phenotypes, which are normally present in small numbers within human tissues. Similarly, cultured normal human mammary epithelial cell lines are enriched for rare basal and mesenchymal phenotypes that represent a minor fraction of cells within reduction mammoplasty tissues. Furthermore, although normal human mammary epithelial cell lines exhibit features of bi-potent progenitor cells they are unable to differentiate into mature luminal breast epithelial cells under standard culture conditions.

Conclusions

As a group breast cancer cell lines represent the heterogeneity of human breast tumors, but individually they exhibit increased lineage-restricted profiles that fall short of truly representing the intratumoral heterogeneity of individual breast tumors. Additionally, normal human mammary epithelial cell lines fail to retain much of the cellular diversity found in human breast tissues and are enriched for differentiation states that are a minority in breast tissues, although they do exhibit features of bi-potent basal progenitor cells. These findings suggest that collections of cell lines representing multiple cell types can be used to model the cellular heterogeneity of tissues.
Zusatzmaterial
Additional file 1: Morphology and surface markers EpCAM, CD24, and CD49f classify breast cancer cell lines into distinct differentiation states. Human Luminal breast cancer cell lines can be classified into Luminal 1 or Luminal 2 cell lines based on morphology in tissue culture (left panels, original magnification: 100×) and by expression of EpCAM, CD24 and CD49f cell surface markers (dot plots, right panels). Cell lines were stained for EpCAM, CD24, and CD49f and quantified by flow cytometry as described in Materials and methods. (PDF 423 KB)
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Additional file 2: Morphology and surface markers EpCAM, CD24, and CD49f classify breast cancer cell lines into distinct differentiation states. Human Basal breast cancer cell lines can be classified into Basal or Mesenchymal cell lines based on morphology in tissue culture (left panels, original magnification: 100×) and by expression of EpCAM, CD24 and CD49f cell surface markers (dot plots, right panels). Cell lines were stained for EpCAM, CD24, and CD49f and quantified by flow cytometry as described in Materials and Methods. (PDF 442 KB)
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Additional file 3: CD44 + /CD24 - /EpCAM + cells are variable across a panel of cultured human breast cell lines. Human breast cancer cell lines were stained for EpCAM, CD24, and CD44 and quantified by flow cytometry as described in Materials and Methods. Cell staining CD44+/CD24- (upper left quadrant, dot plots) were analyzed for the percentage of EpCAM+ cells, which is shown in the histogram to the right of the dot plots. The percentage of CD44+/CD24-/EpCAM+ cells is calculated by multiplying the percentage of EpCAM+ cells by the percentage of CD44+/CD24+ cells. (PDF 628 KB)
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Additional file 4: Luminal 1, Luminal 2, Basal, and Mesenchymal cell lines identified by EpCAM, CD24, and CD49f expression were classified on the basis of CK14, CK8/18, ERα, EpCAM, and vimentin expression. Representative immunofluorescent images are from the panel of Luminal 1, Luminal 2, Basal, and Mesenchymal cell lines. Nuclei were counterstained with DAPI (blue). Original magnification: 200×. (PDF 754 KB)
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Additional file 5: Table 1. Molecular and cellular characterization of human breast cell lines. (XLS 31 KB)
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Additional file 6: Table 2. Histopathological characteristics of breast cancer cell line xenografts. (XLS 117 KB)
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Additional file 7: Table 3. In vitro vs. in vivo comparative molecular marker expression of breast cancer cell lines. (XLS 117 KB)
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Authors’ original file for figure 1
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Authors’ original file for figure 2
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Authors’ original file for figure 3
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Authors’ original file for figure 4
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Authors’ original file for figure 5
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Authors’ original file for figure 6
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