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Numb protein expression correlates with a basal-like phenotype and cancer stem cell markers in primary breast cancer

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Abstract

Decreased expression of Numb, resulting in activation of the proto-oncogene Notch1 and reduction in the tumor suppressor p53, has been demonstrated in mammary carcinomas. The aim of this study was to investigate the relationship between Numb protein expression and clinicopathological characteristics, tumor biological subtypes and putative cancer stem cell markers in a well-characterized cohort of primary human breast cancers. Immunohistochemistry was performed on tissue microarrays of primary invasive breast tumors using a polyclonal anti-Numb primary antibody. Of the 241 tumors evaluated, 50 (21%) displayed deficient or reduced Numb immunoreactivity. Retained Numb expression was significantly correlated to estrogen (ER) and progesterone receptor (PR) positivity (P < 0.001 and P = 0.004, respectively). Interestingly, we found that a higher percentage of the tumors with deficient or reduced Numb expression belonged to the triple-negative (ER-/PR-/HER2−) subgroup compared to tumors with retained Numb expression (P = 0.004). Transcriptional profiling of a subset of these tumors linked NOTCH1 and BIRC5, both downstream targets of Numb, to the triple-negative subgroup in an inverse manner. Typically, subgroups characterized by the low expression of Numb expressed higher levels of NOTCH1 and BIRC5 (encoding survivin). We also found deficient expression of Numb in a significantly higher proportion of BRCA1 dependent tumors, which are usually triple-negative, compared to sporadic tumors. The expression of Numb in 14 breast cancer cell lines correlated similarly to their respective molecular subtypes. We further established an inverse correlation between the Numb expression levels and the CD44+/CD24− cancer stem cell phenotype (P = 0.05) in primary tumors. Finally, decreased Numb expression was associated with poorer distant disease-free survival (P = 0.01). Taken together, our results indicate that loss of Numb expression is a marker of tumor aggressiveness, potentially linked to BRCA1 status and a cancer stem cell phenotype in primary breast cancer.

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Acknowledgments

We thank the participating departments of the South Sweden Breast Cancer Group for their contribution of breast cancer samples, Mårten Fernö and Åke Borg for providing TMA sections and Kristina Lövgren and Mats Jönsson for technical assistance. The study was supported by funds from the Swedish Cancer Society, the Swedish Research Council, the G. Nilsson Cancer Foundation, the B. Kamprad Cancer Foundation, the Å. Wiberg Foundation, the University Hospital of Lund Research Foundation and Governmental Funding of Clinical Research within National Health Service. KR and IH were supported by the Swedish Cancer Society.

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Authors and Affiliations

  1. Department of Oncology, Clinical Sciences, Lund University, Barngatan 2B, 221 85, Lund, Sweden

    Karin Rennstam, Nicole McMichael, Pontus Berglund, Gabriella Honeth, Cecilia Hegardt, Pär-Ola Bendahl & Ingrid Hedenfalk

  2. Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, Lund University, 221 85, Lund, Sweden

    Cecilia Hegardt

  3. Department of Surgery, Clinical Sciences, Lund University, 221 85, Lund, Sweden

    Lisa Rydén

  4. Department of Clinical Pathology, Clinical Sciences, Lund University, 221 85, Lund, Sweden

    Lena Luts

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  1. Karin Rennstam
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  2. Nicole McMichael
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Correspondence to Ingrid Hedenfalk.

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Rennstam, K., McMichael, N., Berglund, P. et al. Numb protein expression correlates with a basal-like phenotype and cancer stem cell markers in primary breast cancer. Breast Cancer Res Treat 122, 315–324 (2010). https://doi.org/10.1007/s10549-009-0568-x

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  • DOI: https://doi.org/10.1007/s10549-009-0568-x

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