HER2 mRNA transcript quantitation in breast cancer

K. Meehan, B. Clynick, B. Mirzai, P. Maslen, Jennet Harvey, W. N. Erber

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    Purpose: The human epidermal growth factor receptor 2 (HER2) status in breast cancer is important for prognostic prediction and the determination of optimal treatment. Current methods rely on protein expression, as determined by immunohistochemistry (IHC), as well as gene amplification as determined by in situ hybridisation (ISH). We explored whether quantitative droplet digital PCR (ddPCR) can be used for the detection and absolute quantitation of HER2 mRNA. Methods: Digital droplet PCR (ddPCR) was performed for HER2 mRNA on 178 formalin-fixed paraffin-embedded (FFPE) breast cancer specimens. HER2 positive, equivocal and negative cases as defined by standard criteria were included and both core biopsies and tissue sections were assessed. Results: HER2 positive cases contained significantly higher levels of HER2 mRNA (169–1,000,000 copies/µl) by ddPCR compared with equivocal (112–139 copies/µl, p = 0.025) and negative cases (6.2–644 copies/µl. p < 0.001). A continuum of transcript quantity was observed but a cutoff of 490 copies/µl distinguished between HER2 positive and negative cases. Results were consistent between core biopsy and tissue sections. Conclusions: ddPCR can be used to quantify HER2 mRNA transcripts in FFPE breast cancer specimens. Our results highlight the potential of ddPCR on FFPE tissue to be used to accurately quantify HER2 transcripts. Validation in large cohorts will be required to determine a clinically applicable cutoff.

    Original languageEnglish
    Pages (from-to)606-615
    Number of pages10
    JournalClinical and Translational Oncology
    DOIs
    Publication statusPublished - 11 Nov 2016

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    Breast Neoplasms
    Messenger RNA
    Polymerase Chain Reaction
    Paraffin
    Formaldehyde
    Biopsy
    human ERBB2 protein
    Gene Amplification
    In Situ Hybridization
    Immunohistochemistry
    Proteins

    Cite this

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    title = "HER2 mRNA transcript quantitation in breast cancer",
    abstract = "Purpose: The human epidermal growth factor receptor 2 (HER2) status in breast cancer is important for prognostic prediction and the determination of optimal treatment. Current methods rely on protein expression, as determined by immunohistochemistry (IHC), as well as gene amplification as determined by in situ hybridisation (ISH). We explored whether quantitative droplet digital PCR (ddPCR) can be used for the detection and absolute quantitation of HER2 mRNA. Methods: Digital droplet PCR (ddPCR) was performed for HER2 mRNA on 178 formalin-fixed paraffin-embedded (FFPE) breast cancer specimens. HER2 positive, equivocal and negative cases as defined by standard criteria were included and both core biopsies and tissue sections were assessed. Results: HER2 positive cases contained significantly higher levels of HER2 mRNA (169–1,000,000 copies/µl) by ddPCR compared with equivocal (112–139 copies/µl, p = 0.025) and negative cases (6.2–644 copies/µl. p < 0.001). A continuum of transcript quantity was observed but a cutoff of 490 copies/µl distinguished between HER2 positive and negative cases. Results were consistent between core biopsy and tissue sections. Conclusions: ddPCR can be used to quantify HER2 mRNA transcripts in FFPE breast cancer specimens. Our results highlight the potential of ddPCR on FFPE tissue to be used to accurately quantify HER2 transcripts. Validation in large cohorts will be required to determine a clinically applicable cutoff.",
    keywords = "Breast cancer, Digital droplet PCR, Human epidermal growth factor receptor",
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    HER2 mRNA transcript quantitation in breast cancer. / Meehan, K.; Clynick, B.; Mirzai, B.; Maslen, P.; Harvey, Jennet; Erber, W. N.

    In: Clinical and Translational Oncology, 11.11.2016, p. 606-615.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - HER2 mRNA transcript quantitation in breast cancer

    AU - Meehan, K.

    AU - Clynick, B.

    AU - Mirzai, B.

    AU - Maslen, P.

    AU - Harvey, Jennet

    AU - Erber, W. N.

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    N2 - Purpose: The human epidermal growth factor receptor 2 (HER2) status in breast cancer is important for prognostic prediction and the determination of optimal treatment. Current methods rely on protein expression, as determined by immunohistochemistry (IHC), as well as gene amplification as determined by in situ hybridisation (ISH). We explored whether quantitative droplet digital PCR (ddPCR) can be used for the detection and absolute quantitation of HER2 mRNA. Methods: Digital droplet PCR (ddPCR) was performed for HER2 mRNA on 178 formalin-fixed paraffin-embedded (FFPE) breast cancer specimens. HER2 positive, equivocal and negative cases as defined by standard criteria were included and both core biopsies and tissue sections were assessed. Results: HER2 positive cases contained significantly higher levels of HER2 mRNA (169–1,000,000 copies/µl) by ddPCR compared with equivocal (112–139 copies/µl, p = 0.025) and negative cases (6.2–644 copies/µl. p < 0.001). A continuum of transcript quantity was observed but a cutoff of 490 copies/µl distinguished between HER2 positive and negative cases. Results were consistent between core biopsy and tissue sections. Conclusions: ddPCR can be used to quantify HER2 mRNA transcripts in FFPE breast cancer specimens. Our results highlight the potential of ddPCR on FFPE tissue to be used to accurately quantify HER2 transcripts. Validation in large cohorts will be required to determine a clinically applicable cutoff.

    AB - Purpose: The human epidermal growth factor receptor 2 (HER2) status in breast cancer is important for prognostic prediction and the determination of optimal treatment. Current methods rely on protein expression, as determined by immunohistochemistry (IHC), as well as gene amplification as determined by in situ hybridisation (ISH). We explored whether quantitative droplet digital PCR (ddPCR) can be used for the detection and absolute quantitation of HER2 mRNA. Methods: Digital droplet PCR (ddPCR) was performed for HER2 mRNA on 178 formalin-fixed paraffin-embedded (FFPE) breast cancer specimens. HER2 positive, equivocal and negative cases as defined by standard criteria were included and both core biopsies and tissue sections were assessed. Results: HER2 positive cases contained significantly higher levels of HER2 mRNA (169–1,000,000 copies/µl) by ddPCR compared with equivocal (112–139 copies/µl, p = 0.025) and negative cases (6.2–644 copies/µl. p < 0.001). A continuum of transcript quantity was observed but a cutoff of 490 copies/µl distinguished between HER2 positive and negative cases. Results were consistent between core biopsy and tissue sections. Conclusions: ddPCR can be used to quantify HER2 mRNA transcripts in FFPE breast cancer specimens. Our results highlight the potential of ddPCR on FFPE tissue to be used to accurately quantify HER2 transcripts. Validation in large cohorts will be required to determine a clinically applicable cutoff.

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