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Cytokeratin 5 positive cells represent a steroid receptor negative and therapy resistant subpopulation in luminal breast cancers

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Abstract

A majority of breast cancers are estrogen receptor (ER) positive and have a luminal epithelial phenotype. However, these ER+ tumors often contain heterogeneous subpopulations of ER− tumor cells. We previously identified a population of cytokeratin 5 (CK5) positive cells within ER+ and progesterone receptor positive (PR+) tumors that is both ER−PR− and CD44+, a marker of breast tumor-initiating cells (TICs). These CK5+ cells have properties of TICs in luminal tumor xenografts, and we speculated that they are more resistant to chemo- and anti-ER-targeted therapies than their ER+ neighbors. To test this, we used ER+PR+ T47D and MCF7 breast cancer cells. CK5+ cells had lower proliferative indices than CK5− cells, were less sensitive to 5-fluorouracil and docetaxel, and cultures became enriched for CK5+ cells after treatments. CK5+ cells were less prone to drug-induced apoptosis than CK5− cells. In cells treated with 17β-estradiol (E) plus anti-estrogens tamoxifen or fulvestrant, ER protein levels decreased, and CK5 protein levels increased, compared to controls treated with E alone. In ER+ tumors from patients treated with neoadjuvant endocrine therapies ER gene expression decreased, and CK5 gene expression increased in post compared to pre-treatment tumors. The number of CK5+ cells in tumors also increased in post- compared to pre-treatment tumors. We conclude that an ER−PR−CK5+ subpopulation found in many luminal tumors is resistant to standard endocrine and chemotherapies, relative to the majority ER+PR+CK5− cells. Compounds that effectively target these cells are needed to improve outcome in luminal breast cancers.

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Abbreviations

AI:

Aromatase inhibitor

ALDH1:

Aldehyde dehydrogenase 1

BrdU:

5-Bromo-2-deoxyuridine

CK:

Cytokeratin

2D:

Two-dimensional

3D:

Three dimensional

DAPI:

4′,6-Diamidino-2-phenylindole

Dx:

Docetaxel

EGFR:

Epidermal growth factor receptor

ER:

Estrogen receptor

E:

17β-estradiol

ESA:

Epithelial specific antigen

EWD:

Estrogen withdrawal

FBS:

Fetal bovine serum

5-FU:

5-Fluorouracil

HER2:

Human epidermal growth factor receptor 2

ICC:

Immunocytochemistry

IHC:

Immunohistochemistry

Lin:

Lineage

MPA:

Medroxyprogesterone acetate

PR:

Progesterone receptor

SOC:

Sodium deoxycholate

Tam:

4-Hydroxytamoxifen

TIC:

Tumor initiating cell

TNP:

Triple negative phenotype

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Acknowledgments

This work was supported by grants from the American Society of Clinical Oncology YIA-PN200810-161 (P. Kabos), National Institutes of Health 1 F32 CA142096-01 (J.M. Haughian), Martha Cannon Dear Professorship (A. Elias), National Institutes of Health R01 CA26869 (K.B. Horwitz), Breast Cancer Research Foundation (K.B. Horwitz), the National Foundation for Cancer Research (K.B. Horwitz), The Avon Foundation (K.B. Horwitz), The Avon Foundation (C.A. Sartorius), and the University of Colorado Cancer Center (C.A. Sartorius).

Conflicts of interest

The authors have no conflicts of interest to disclose.

Author information

Authors and Affiliations

  1. Division of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO, 80045, USA

    Peter Kabos & Anthony Elias

  2. Division of Endocrinology, Metabolism, and Diabetes, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO, 80045, USA

    James M. Haughian, Xinshuo Wang, Wendy W. Dye, Kathryn B. Horwitz & Carol A. Sartorius

  3. Department of Surgery, Department of Medicine, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO, 80045, USA

    Christina Finlayson

  4. Department of Pathology, University of Colorado Anschutz Medical Campus, 12801 E 17th Ave, Aurora, CO, 80045, USA

    Kathryn B. Horwitz

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  1. Peter Kabos
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Correspondence to Carol A. Sartorius.

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Kabos, P., Haughian, J.M., Wang, X. et al. Cytokeratin 5 positive cells represent a steroid receptor negative and therapy resistant subpopulation in luminal breast cancers. Breast Cancer Res Treat 128, 45–55 (2011). https://doi.org/10.1007/s10549-010-1078-6

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  • DOI: https://doi.org/10.1007/s10549-010-1078-6

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