Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer

Willemijne A M E Schrijver, Petra van der Groep, Laurien DC Hoefnagel, Natalie D. ter Hoeve, Ton Peeters, Cathy B. Moelans, Paul J. van Diest

Research output: Contribution to journalArticle

Abstract

Distant breast cancer metastases are nowadays routinely biopsied to reassess receptor status and to isolate DNA for sequencing of druggable targets. Bone metastases are the most frequent subgroup. Decalcification procedures may negatively affect antigenicity and DNA quality. We therefore evaluated the effect of several decalcification procedures on receptor status and DNA/RNA quality. In 23 prospectively collected breast tumors, we compared ERα, PR and HER2 status by immunohistochemistry in (non-decalcified) tissue routinely processed for diagnostic purposes and in parallel tissue decalcified in Christensen’s buffer with and without microwave, EDTA and Formical-4. Furthermore, HER2 fluorescence in situ hybridization and DNA/RNA quantity and quality were assessed. We found that the percentage of ERα-positive cells were on average lower in EDTA (P=0.049) and Formical-4 (P=0.047) treated cases, compared with controls, and PR expression showed decreased antigenicity after Christensen’s buffer treatment (P=0.041). Overall, a good concordance (weighted kappa) was seen for ERα, PR and HER2 immunohistochemistry when comparing the non-decalcified control tissues with the decalcified tissues. For two patients (9%), there was a potential influence on therapeutic decision making with regard to hormonal therapy or HER2-targeted therapy. HER2 fluorescence in situ hybridization interpretation was seriously hampered by Christensen’s buffer and Formical-4, and DNA/RNA quantity and quality were decreased after all four decalcification procedures. Validation on paired primary breast tumor specimens and EDTA-treated bone metastases showed that immunohistochemistry and fluorescence in situ hybridization were well assessable and DNA and RNA yield and quality were sufficient. With this, we conclude that common decalcification procedures have only a modest negative influence on hormone and HER2 receptor immunohistochemistry in breast cancer. However, they may seriously affect DNA/RNA-based diagnostic procedures. Overall, EDTA-based decalcification is therefore to be preferred as it best allows fluorescence in situ hybridization and DNA/RNA isolation.Modern Pathology advance online publication, 26 August 2016; doi:10.1038/modpathol.2016.116.

Original languageEnglish (US)
JournalModern Pathology
DOIs
StateAccepted/In press - Aug 26 2016
Externally publishedYes

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Molecular Pathology
Immunohistochemistry
RNA
Breast Neoplasms
Fluorescence In Situ Hybridization
Edetic Acid
DNA
Buffers
Neoplasm Metastasis
Bone and Bones
Therapeutics
Microwaves
DNA Sequence Analysis
Publications
Decision Making
Hormones
Pathology

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Medicine(all)

Cite this

Schrijver, W. A. M. E., van der Groep, P., Hoefnagel, L. DC., ter Hoeve, N. D., Peeters, T., Moelans, C. B., & van Diest, P. J. (Accepted/In press). Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer. Modern Pathology. https://doi.org/10.1038/modpathol.2016.116

Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer. / Schrijver, Willemijne A M E; van der Groep, Petra; Hoefnagel, Laurien DC; ter Hoeve, Natalie D.; Peeters, Ton; Moelans, Cathy B.; van Diest, Paul J.

In: Modern Pathology, 26.08.2016.

Research output: Contribution to journalArticle

Schrijver, WAME, van der Groep, P, Hoefnagel, LDC, ter Hoeve, ND, Peeters, T, Moelans, CB & van Diest, PJ 2016, 'Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer', Modern Pathology. https://doi.org/10.1038/modpathol.2016.116
Schrijver WAME, van der Groep P, Hoefnagel LDC, ter Hoeve ND, Peeters T, Moelans CB et al. Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer. Modern Pathology. 2016 Aug 26. https://doi.org/10.1038/modpathol.2016.116
Schrijver, Willemijne A M E ; van der Groep, Petra ; Hoefnagel, Laurien DC ; ter Hoeve, Natalie D. ; Peeters, Ton ; Moelans, Cathy B. ; van Diest, Paul J. / Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer. In: Modern Pathology. 2016.
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