Quantitative RT-PCR assay of HER2 mRNA expression in formalin-fixed and paraffin-embedded breast cancer tissues

Sangjung Park, Hye Young Wang, Sunghyun Kim, Sungwoo Ahn, Dongsup Lee, Yoonjung Cho, Kwang Hwa Park, Dongju Jung, Seung Il Kim, Hyeyoung Lee

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Detection of human epidermal growth factor receptor 2 gene (HER2, also known as erbB2) expression is a preparatory process to decide a treatment strategy for breast cancer patients. 20-30% of breast cancer patients have HER2 overexpression, and they usually show poor recovery rate. For detection of HER2 expression, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) methods are conventionally used. Although these methods are accurate and reliable, their time-consuming process and high cost need a concise method with high sensitivity and accuracy. As a complementary method to the current IHC/FISH standard techniques, PCR-based methods have been developed. Here we employed a quantitative PCR method to detect HER2 expression in one hundred ninety nine formalin-fixed and paraffin-embedded (FFPE) breast cancer tissue samples from the patients treated over two years at the Yonsei University Severance Hospital, Republic of Korea. Relative expression of HER2 mRNA in the FFPE samples was analyzed using a quantitative RT-PCR (RT-qPCR) method and the obtained HER2 expression levels were compared with those from IHC/FISH methods. Our results show that the RT-qPCR method was highly concordant with IHC/FISH methods for detecting HER2 expression. Overall sensitivity and specificity of the BrightGen HER2 RT-qDx assay kit (Syantra, Calgary, Canada), which is a kit we used for RT-qPCR analyses, were 93.0% and 89.8% (P < 0.0001), respectively. The diagnostic cut-off value of HER2 RT-qDx for the clinical samples was determined by likelihood ratio, among which the highest likelihood ratio of relative HER2 mRNA levels was over 105.5 (AUC = 0.9466) with the highest sensitivity and specificity. Our study indicates that quantification of HER2 mRNA expression with the RT-qPCR could be an alternative method of conventional IHC/FISH methods.

Original languageEnglish
Pages (from-to)6752-6759
Number of pages8
JournalInternational Journal of Clinical and Experimental Pathology
Volume7
Issue number10
Publication statusPublished - 2014 Jan 1

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Paraffin
Formaldehyde
Breast Neoplasms
Messenger RNA
Fluorescence In Situ Hybridization
Immunohistochemistry
erbB-1 Genes
Sensitivity and Specificity
Polymerase Chain Reaction
Republic of Korea
Area Under Curve
Canada
Costs and Cost Analysis

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Histology

Cite this

Park, Sangjung ; Wang, Hye Young ; Kim, Sunghyun ; Ahn, Sungwoo ; Lee, Dongsup ; Cho, Yoonjung ; Park, Kwang Hwa ; Jung, Dongju ; Kim, Seung Il ; Lee, Hyeyoung. / Quantitative RT-PCR assay of HER2 mRNA expression in formalin-fixed and paraffin-embedded breast cancer tissues. In: International Journal of Clinical and Experimental Pathology. 2014 ; Vol. 7, No. 10. pp. 6752-6759.
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title = "Quantitative RT-PCR assay of HER2 mRNA expression in formalin-fixed and paraffin-embedded breast cancer tissues",
abstract = "Detection of human epidermal growth factor receptor 2 gene (HER2, also known as erbB2) expression is a preparatory process to decide a treatment strategy for breast cancer patients. 20-30{\%} of breast cancer patients have HER2 overexpression, and they usually show poor recovery rate. For detection of HER2 expression, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) methods are conventionally used. Although these methods are accurate and reliable, their time-consuming process and high cost need a concise method with high sensitivity and accuracy. As a complementary method to the current IHC/FISH standard techniques, PCR-based methods have been developed. Here we employed a quantitative PCR method to detect HER2 expression in one hundred ninety nine formalin-fixed and paraffin-embedded (FFPE) breast cancer tissue samples from the patients treated over two years at the Yonsei University Severance Hospital, Republic of Korea. Relative expression of HER2 mRNA in the FFPE samples was analyzed using a quantitative RT-PCR (RT-qPCR) method and the obtained HER2 expression levels were compared with those from IHC/FISH methods. Our results show that the RT-qPCR method was highly concordant with IHC/FISH methods for detecting HER2 expression. Overall sensitivity and specificity of the BrightGen HER2 RT-qDx assay kit (Syantra, Calgary, Canada), which is a kit we used for RT-qPCR analyses, were 93.0{\%} and 89.8{\%} (P < 0.0001), respectively. The diagnostic cut-off value of HER2 RT-qDx for the clinical samples was determined by likelihood ratio, among which the highest likelihood ratio of relative HER2 mRNA levels was over 105.5 (AUC = 0.9466) with the highest sensitivity and specificity. Our study indicates that quantification of HER2 mRNA expression with the RT-qPCR could be an alternative method of conventional IHC/FISH methods.",
author = "Sangjung Park and Wang, {Hye Young} and Sunghyun Kim and Sungwoo Ahn and Dongsup Lee and Yoonjung Cho and Park, {Kwang Hwa} and Dongju Jung and Kim, {Seung Il} and Hyeyoung Lee",
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Park, S, Wang, HY, Kim, S, Ahn, S, Lee, D, Cho, Y, Park, KH, Jung, D, Kim, SI & Lee, H 2014, 'Quantitative RT-PCR assay of HER2 mRNA expression in formalin-fixed and paraffin-embedded breast cancer tissues', International Journal of Clinical and Experimental Pathology, vol. 7, no. 10, pp. 6752-6759.

Quantitative RT-PCR assay of HER2 mRNA expression in formalin-fixed and paraffin-embedded breast cancer tissues. / Park, Sangjung; Wang, Hye Young; Kim, Sunghyun; Ahn, Sungwoo; Lee, Dongsup; Cho, Yoonjung; Park, Kwang Hwa; Jung, Dongju; Kim, Seung Il; Lee, Hyeyoung.

In: International Journal of Clinical and Experimental Pathology, Vol. 7, No. 10, 01.01.2014, p. 6752-6759.

Research output: Contribution to journalArticle

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T1 - Quantitative RT-PCR assay of HER2 mRNA expression in formalin-fixed and paraffin-embedded breast cancer tissues

AU - Park, Sangjung

AU - Wang, Hye Young

AU - Kim, Sunghyun

AU - Ahn, Sungwoo

AU - Lee, Dongsup

AU - Cho, Yoonjung

AU - Park, Kwang Hwa

AU - Jung, Dongju

AU - Kim, Seung Il

AU - Lee, Hyeyoung

PY - 2014/1/1

Y1 - 2014/1/1

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