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Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues
Breast cancers with amplification and overexpression of human epithelial growth factor receptor 2 (HER2) are associated with poor prognosis, and targeted for anti-HER2 therapy. Immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) are currently the recommended methods to asses HER...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer US
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6182624/ https://www.ncbi.nlm.nih.gov/pubmed/30284063 http://dx.doi.org/10.1007/s12032-018-1210-8 |
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author | Tantiwetrueangdet, Anchalee Panvichian, Ravat Wongwaisayawan, Sansanee Sueangoen, Natthaporn Lertsithichai, Panuwat |
author_facet | Tantiwetrueangdet, Anchalee Panvichian, Ravat Wongwaisayawan, Sansanee Sueangoen, Natthaporn Lertsithichai, Panuwat |
author_sort | Tantiwetrueangdet, Anchalee |
collection | PubMed |
description | Breast cancers with amplification and overexpression of human epithelial growth factor receptor 2 (HER2) are associated with poor prognosis, and targeted for anti-HER2 therapy. Immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) are currently the recommended methods to asses HER2 overexpression/amplification. Droplet digital PCR (ddPCR), a highly accurate method to quantify DNA copy number, is potentially a robust alternative for HER2 diagnostics. In the FISH assay and most of previous ddPCR reports, chromosome 17 centromere (CEP17) has been used as the reference control to determine HER2/CEP17 ratio. Nevertheless, miss-classification could occur when HER2 is co-amplified with CEP17. To avoid this inherent defect, in the present study, we employed ddPCR assay using the human eukaryotic translation initiation factor 2C1 (EIF2C1) gene located at chromosome 1p34.3 as the reference control to quantify HER2 copy number in 31 frozen breast cancer tissues. HER2 status of these samples had been determined by FISH and classified as HER2-amplified and HER2-non-amplified breast cancers. The results showed that HER2 determined by ddPCR using HER2/EIF2C1 ratio was in good concordance with HER2 determined by FISH using HER2/CEP17 ratio, the concordance rate 87.1% (27/31), Kappa = 0.719. The sensitivity and specificity of ddPCR assay was 90% (9/10) and 85.7% (18/21), respectively. The median HER2/EIF2C1 copy number ratio in HER2-amplified cancers (6.55, range 1.3–17.3) was significantly higher than in HER2-non-amplified cancers (1.05, range 0.6–3.6, p < 0.001). This study demonstrated that ddPCR using HER2/EIF2C1 ratio could accurately assess HER2 status in frozen breast cancer tissues. Thus, our findings warrant further studies into breast cancer with HER2-equivocal by IHC/FISH. |
format | Online Article Text |
id | pubmed-6182624 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Springer US |
record_format | MEDLINE/PubMed |
spelling | pubmed-61826242018-10-24 Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues Tantiwetrueangdet, Anchalee Panvichian, Ravat Wongwaisayawan, Sansanee Sueangoen, Natthaporn Lertsithichai, Panuwat Med Oncol Original Paper Breast cancers with amplification and overexpression of human epithelial growth factor receptor 2 (HER2) are associated with poor prognosis, and targeted for anti-HER2 therapy. Immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) are currently the recommended methods to asses HER2 overexpression/amplification. Droplet digital PCR (ddPCR), a highly accurate method to quantify DNA copy number, is potentially a robust alternative for HER2 diagnostics. In the FISH assay and most of previous ddPCR reports, chromosome 17 centromere (CEP17) has been used as the reference control to determine HER2/CEP17 ratio. Nevertheless, miss-classification could occur when HER2 is co-amplified with CEP17. To avoid this inherent defect, in the present study, we employed ddPCR assay using the human eukaryotic translation initiation factor 2C1 (EIF2C1) gene located at chromosome 1p34.3 as the reference control to quantify HER2 copy number in 31 frozen breast cancer tissues. HER2 status of these samples had been determined by FISH and classified as HER2-amplified and HER2-non-amplified breast cancers. The results showed that HER2 determined by ddPCR using HER2/EIF2C1 ratio was in good concordance with HER2 determined by FISH using HER2/CEP17 ratio, the concordance rate 87.1% (27/31), Kappa = 0.719. The sensitivity and specificity of ddPCR assay was 90% (9/10) and 85.7% (18/21), respectively. The median HER2/EIF2C1 copy number ratio in HER2-amplified cancers (6.55, range 1.3–17.3) was significantly higher than in HER2-non-amplified cancers (1.05, range 0.6–3.6, p < 0.001). This study demonstrated that ddPCR using HER2/EIF2C1 ratio could accurately assess HER2 status in frozen breast cancer tissues. Thus, our findings warrant further studies into breast cancer with HER2-equivocal by IHC/FISH. Springer US 2018-10-03 2018 /pmc/articles/PMC6182624/ /pubmed/30284063 http://dx.doi.org/10.1007/s12032-018-1210-8 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Paper Tantiwetrueangdet, Anchalee Panvichian, Ravat Wongwaisayawan, Sansanee Sueangoen, Natthaporn Lertsithichai, Panuwat Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues |
title | Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues |
title_full | Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues |
title_fullStr | Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues |
title_full_unstemmed | Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues |
title_short | Droplet digital PCR using HER2/EIF2C1 ratio for detection of HER2 amplification in breast cancer tissues |
title_sort | droplet digital pcr using her2/eif2c1 ratio for detection of her2 amplification in breast cancer tissues |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6182624/ https://www.ncbi.nlm.nih.gov/pubmed/30284063 http://dx.doi.org/10.1007/s12032-018-1210-8 |
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