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Clinical Usefulness of a One-Tube Nested Reverse Transcription Quantitative Polymerase Chain Reaction Assay for Evaluating Human Epidermal Growth Factor Receptor 2 mRNA Overexpression in Formalin-Fixed and Paraffin-Embedded Breast Cancer Tissue Samples

 Wang H.a  ;  Ahn S.b  ;  Park S.b  ;  Kim S.c  ;  Lee H.b 
 PATHOBIOLOGY, Vol.84(2) : 57-70, 2017 
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Issue Date
Biomarkers, Tumor ; Breast/metabolism* ; Breast/pathology ; Breast Neoplasms/genetics* ; Breast Neoplasms/metabolism ; Breast Neoplasms/pathology ; Cell Line, Tumor ; Female ; Formaldehyde ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Paraffin Embedding ; Real-Time Polymerase Chain Reaction ; Receptor, ErbB-2/genetics* ; Receptor, ErbB-2/metabolism ; Sensitivity and Specificity ; Tissue Fixation
Human epidermal growth factor receptor 2 overexpression ; Molecular diagnosis ; Immunohistochemistry ; Fluorescence in situ hybridization ; One-tube nested reverse transcription quantitative polymerase chain reaction ; Breast cancer
BACKGROUND: Currently, the two main methods used to analyze human epidermal growth factor receptor 2 (HER2) amplification or overexpression have a limited accuracy and high costs. These limitations can be overcome by the development of complementary quantitative methods. METHODS: In this study, we analyzed HER2 mRNA expression in clinical formalin-fixed and paraffin-embedded (FFPE) samples using a one-tube nested reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay. We measured expression relative to 3 reference genes and compared the results to those obtained by conventional immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) assays with 226 FFPE breast cancer tissue samples. RESULTS: The one-tube nested RT-qPCR assay proved to be highly sensitive and specific based on comparisons with IHC (96.9 and 97.7%, respectively) and FISH (92.4 and 92.9%, respectively) obtained with the validation set. Comparisons with clinicopathological data revealed significant associations between HER2 overexpression and TNM stage (p < 0.01), histological type (p < 0.01), ER status (p < 0.001), PR status (p < 0.05), HER2 status (p < 0.001), and molecular subtypes (p < 0.001). CONCLUSION: Based on these findings, our one-tube nested RT-qPCR assay is a potentially useful and complementary screening tool for the detection of HER2 mRNA overexpression.
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1. College of Medicine (의과대학) > Dept. of Surgery (외과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Seung Il(김승일)
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