Novel cell culture technique for primary ductal carcinoma in situ: Role of notch and epidermal growth Factor Receptor Signaling Pathways

Gillian Farnie, Robert B. Clarke, Katherine Spence, Natasha Pinnock, Keith Brennan, Neil G. Anderson, Nigel J. Bundred

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Background: The epidermal growth factor receptor (EGFR) and Notch signaling pathways have been implicated in self-renewal of normal breast stem cells. We investigated the involvement of these signaling pathways in ductal carcinoma in situ (DCIS) of the breast. Methods: Samples of normal breast tissue (n = 15), pure DCIS tissue of varying grades (n = 35), and DCIS tissue surrounding an invasive cancer (n = 7) were used for nonadherent (i.e., mammosphere) culture. Mammosphere cultures were treated at day 0 with gefitinib (an EGFR inhibitor), DAPT (N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butyl ester) (a γ-secretase inhibitor), or Notch 4-neutralizing antibody. Mammosphere-forming efficiency (MFE) was calculated by dividing the number of mammospheres of 60 μm or more formed by the number of single cells seeded and is expressed as a percentage. The Notch 1 intracellular domain (NICD) was detected immunohistochemically in paraffin-embedded DCIS tissue from 50 patients with at least 60 months of follow-up. All statistical tests were two-sided. Results: DCIS had agreater MFE than normal breast tissue (1.5% versus 0.5%, difference = 1%, 95% confidence interval [CI] 0.62% to 1.25%,P
    Original languageEnglish
    Pages (from-to)616-627
    Number of pages11
    JournalJournal of the National Cancer Institute
    Volume99
    Issue number8
    DOIs
    Publication statusPublished - 18 Apr 2007

    Keywords

    • cytology: Breast
    • pathology: Breast Neoplasms
    • pathology: Carcinoma in Situ
    • pathology: Carcinoma, Intraductal, Noninfiltrating
    • Cell Adhesion
    • methods: Cell Culture Techniques
    • Female
    • Humans
    • Neoplasm Invasiveness
    • physiology: Receptor, Epidermal Growth Factor
    • physiology: Receptors, Notch
    • Recurrence
    • Tumor Cells, Cultured

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