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CD44 is overexpressed in basal-like breast cancers but is not a driver of 11p13 amplification

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Abstract

Overexpression and alternative splicing of CD44 have been implicated in tumour progression. Here we describe the identification of a high level amplification of human 11p13, encompassing the CD44 gene, in primary breast cancers and cell lines and test whether CD44 acts as the driver of this amplicon. aCGH analysis revealed 11p13 amplification in 3% (3/100) of primary breast carcinomas and in two cell lines. The minimal region of amplification was 34.38–37.62 Mb. Amplification was confirmed by dual-colour FISH in these cell lines and further validated by CISH in an independent tumour cohort. CD44 expression in primary breast cancers was significantly associated with features of basal-like breast cancer. Detection of CD44 expression in breast cancer cell lines confirmed moderate to high expression in basal-like cell lines and minimal expression in luminal cell lines. In both, primary breast cancers and cell lines, 11p13 amplification was associated with high levels of CD44 mRNA expression. CD44 alternative splicing was detected in four of nine cell lines and in tumour samples, irrespective of the amplification status. RNAi mediated knock down of CD44 failed to reveal an increased dependence on CD44 expression for proliferation or survival in amplified cell lines. Given that expression of CD44 is not an absolute requirement for the survival of cells harbouring CD44 gene amplification, CD44 is unlikely to be a driver of the 11p13 amplicon.

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Abbreviations

aCGH:

Array chromosomal genomic hybridisation

CISH:

Chromogenic in situ hybridisation

Ck:

Cytokeratin

ER:

Oestrogen receptor

EGFR:

Epidermal growth factor receptor

FISH:

Fluorescent in situ hybridisation

PR:

Progesterone receptor

qRT-PCR:

Quantitative real-time polymerase chain reaction

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Acknowledgments

This work was funded by Breakthrough Breast Cancer grants to JSR-F and CMI. We acknowledge NHS funding to the NIHR Biomedical Research Centre. We thank Kay Savage and Suzanne Parry (Breakthrough Histopathology Laboratory, The Institute of Cancer Research) who performed the TMA immunohistochemical staining.

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Authors and Affiliations

  1. Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, 237 Fulham Road, London, SW3 6JB, UK

    Pamela Klingbeil, Rachael Natrajan, Gemma Everitt, Radost Vatcheva, Caterina Marchio, Jorge S. Reis-Filho & Clare M. Isacke

  2. Servicio de Anatomia Patologica, Hospital Virgen del Rocío, Seville, Spain

    Jose Palacios

  3. Institute of Pathology, Paderborn, Germany

    Horst Buerger

Authors
  1. Pamela Klingbeil
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  2. Rachael Natrajan
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  3. Gemma Everitt
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  4. Radost Vatcheva
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  5. Caterina Marchio
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  6. Jose Palacios
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  7. Horst Buerger
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  8. Jorge S. Reis-Filho
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  9. Clare M. Isacke
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Corresponding authors

Correspondence to Jorge S. Reis-Filho or Clare M. Isacke.

Additional information

Pamela Klingbeil and Rachael Natrajan have contributed equally to this work. RN and RV performed aCGH and data analysis and RV performed FISH and CISH. CM and JSR-F interpreted the immunohistochemical and in situ hybridisation experiments. JP and HB provided the breast cancer samples for aCGH. PK and GE performed all the cell based assays. CMI and JSR-F designed the experiments. PK, RN, JSR-F and CMI wrote the manuscript.

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Klingbeil, P., Natrajan, R., Everitt, G. et al. CD44 is overexpressed in basal-like breast cancers but is not a driver of 11p13 amplification. Breast Cancer Res Treat 120, 95–109 (2010). https://doi.org/10.1007/s10549-009-0380-7

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