HER-2/neu amplification testing in breast cancer by multiplex ligation-dependent probe amplification in comparison with immunohistochemistry and in situ hybridization.

Content Provider	Europe PMC
Author	Moelans, Cathy B. de Weger, Roel A. van Blokland, Marja T.M. Ezendam, Chantal Elshof, Sabrina Tilanus, Marcel G. J. van Diest, Paul J.
Copyright Year	2009
Abstract	Background: Assessment of HER-2/neu status in invasive breast cancer is crucial to establish eligibility for trastuzumab and taxane based chemotherapy. Next to immunohistochemistry (IHC) to evaluate protein overexpression, a second line gene amplification test is required for cases with equivocal protein expression. This study aimed to validate a new PCR based test, called Multiplex Ligation-dependent Probe Amplification (MLPA), as a simple and quick method to assess HER-2/neu gene amplification status in invasive breast cancer.Methods: MPLA results were compared with gene amplification status assessed by fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) as gold standard, and with protein overexpression by IHC in 518 breast carcinoma patients.Results: About 10% of cases overexpressed HER-2/neu at the protein level (IHC), and 11% of cases showed gene-amplification by MLPA. A high concordance was found between FISH and CISH, MLPA and IHC, and MLPA and CISH. MLPA showed amplification in 7/36 (19%) of the equivocal IHC 2+ cases. However, of the IHC 0/1+ cases, 6/434 (1.4%) were also amplified by MLPA, and amplification was confirmed in all of these cases by FISH/CISH. On the other hand, one of the 48 (2%) IHC 3+ cases was normal by MLPA and lack of amplification was confirmed by FISH/CISH.Conclusion: MLPA is a fast, accurate and cheap method to detect breast cancer HER-2/neu amplification in small quantities of DNA extracted from paraffin blocks, and thereby a reliable alternative to FISH and CISH.
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ISSN	15705870
Journal	Cellular Oncology : the Official Journal of the International Society for Cellular Oncology [Cell Oncol]
Volume Number	31
PubMed Central reference number	PMC4618800
Issue Number	1
PubMed reference number	19096145
e-ISSN	18758606
DOI	10.3233/clo-2009-0461
Language	English
Publisher	IOS Press
Publisher Date	2009-01-01
Access Restriction	Open
Rights License	Copyright © 2009 Hindawi Publishing Corporation and the authors.
Subject Keyword	HER-2/neu amplification multiplex ligation dependent probe amplification immunohistochem- istry
Content Type	Text
Resource Type	Article
Subject	Cancer Research Molecular Medicine Oncology