PMID- 28039034
OWN - NLM
STAT- MEDLINE
DCOM- 20170206
LR  - 20170206
IS  - 1879-0038 (Electronic)
IS  - 0378-1119 (Linking)
VI  - 605
DP  - 2017 Mar 20
TI  - Development and evaluation of a novel RT-qPCR based test for the quantification 
      of HER2 gene expression in breast cancer.
PG  - 114-122
LID - S0378-1119(16)31027-7 [pii]
LID - 10.1016/j.gene.2016.12.027 [doi]
AB  - Accurate measurement of Human epidermal growth factor receptor (HER2) gene 
      expression is central for breast or stomach cancer therapy orientation and 
      prognosis. The current standards testing methods for HER2 expression are 
      immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). In the 
      current study, we explored the use of quantitative real time reverse 
      transcription-PCR (RT-qPCR) as a potential method for the accurate relative 
      quantification of the HER2 gene using formalin fixed paraffin embedded (FFPE) 
      breast cancer biopsy samples. The main aim of the current study is to measure the 
      level of concordance of RT-qPCR based quantification of HER2 overexpression with 
      both IHC and FISH. Accordingly, an endogenous control gene (ECG) is required for 
      this relative quantification and should ideally be expressed equivalently across 
      tested samples. Stably expressed ECGs have been selected from a panel of seven 
      genes using GenEx V6 software which is based on geNorm and NormFinder and 
      statistical methods. Quantification of HER2 gene expression was performed by our 
      RT-qPCR-based test and compared to the results obtained by both IHC and FISH 
      methods. HER2 gene quantification using RT-qPCR test was normalized using the two 
      ECGs (RPL30 and RPL37A) that were successfully identified and selected from a 
      panel of seven genes as the most stable and reliable ECGs. We evaluated a total 
      of 216 FFPE tissue samples from breast cancer patients. The results obtained with 
      RT-qPCR in the current study were compared to both IHC and FISH data collected 
      for the same patients. In addition to an internal evaluation, an external 
      evaluation of this assay was also performed in a recognized pathology center in 
      Europe (Clinic Barcelona Hospital Universitari, Spain) using 116 FFPE breast 
      cancer tissue samples. The results demonstrated a high concordance between 
      RT-qPCR and either IHC (98%) or FISH (72%) methods. Accordantly, the overall 
      concordance was 85%. To our knowledge, this is the first study using the specific 
      combination of RPL30 and RPL37 as reference genes for an accurate HER2 gene 
      quantification in FFPE biopsy samples. Although further clinical validation 
      regarding evolution and therapeutic response using RT-qPCR for the quantification 
      of HER2 expression are still needed, the present study constitutes definitely a 
      factual element that the RT-qPCR based assay may constitute a valid complementary 
      test to accurately measure HER2 expression for a better treatment orientation.
CI  - Copyright (c) 2017 Elsevier B.V. All rights reserved.
FAU - El Hadi, Hicham
AU  - El Hadi H
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco; 
      Biochemistry and Immunology Laboratory, Faculty of Science, Rabat University 
      Mohamed V, Avenue Ibn Battouta 1014, Rabat, Morocco. Electronic address: 
      h.elhadi@mascir.com.
FAU - Abdellaoui-Maane, Imane
AU  - Abdellaoui-Maane I
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco; 
      Biochemistry and Immunology Laboratory, Faculty of Science, Rabat University 
      Mohamed V, Avenue Ibn Battouta 1014, Rabat, Morocco. Electronic address: 
      i.abdellaoui@mascir.com.
FAU - Kottwitz, Denise
AU  - Kottwitz D
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco. 
      Electronic address: denisekottwitz@web.de.
FAU - El Amrani, Manal
AU  - El Amrani M
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco. 
      Electronic address: dahmani_manale@yahoo.fr.
FAU - Bouchoutrouch, Nadia
AU  - Bouchoutrouch N
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco. 
      Electronic address: n.bouchoutrouch@mascir.com.
FAU - Qmichou, Zineb
AU  - Qmichou Z
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco. 
      Electronic address: z.qmichou@mascir.com.
FAU - Karkouri, Mehdi
AU  - Karkouri M
AD  - Pathology Department, HOPITAL IBN ROCHD, Quartier Des Hopitaux, Casablanca, 
      Morocco. Electronic address: mehdi.karkouri@menara.ma.
FAU - ElAttar, Hicham
AU  - ElAttar H
AD  - Laboratory of Pathology MlyIdriss I, Boulevard MoulayIdriss 1er Quartier Des 
      Hopitaux, Casablanca, Morocco.
FAU - Errihani, Hassan
AU  - Errihani H
AD  - National Institute of Oncology (INO), Avenue Allal El Fassi, Rabat, Morocco.
FAU - Fernandez, Pedro L
AU  - Fernandez PL
AD  - Department of Anatomical Pathology, Hospital Clinic and Institut d'Investigacions 
      Biomediques August Pi iSunyer (IDIBAPS), University of Barcelona, Spain. 
      Electronic address: plfernan@clinic.ub.es.
FAU - Bakri, Youssef
AU  - Bakri Y
AD  - Biochemistry and Immunology Laboratory, Faculty of Science, Rabat University 
      Mohamed V, Avenue Ibn Battouta 1014, Rabat, Morocco. Electronic address: 
      ybakri@gmail.com.
FAU - Sefrioui, Hassan
AU  - Sefrioui H
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco. 
      Electronic address: h.sefrioui@mascir.com.
FAU - Moumen, Abdeladim
AU  - Moumen A
AD  - Medical Biotechnology Center, Moroccan Foundation for Advanced Science Innovation 
      and Research (MASCIR), Rue Mohamed Al JazouliMadinat Al Irfane, Rabat, Morocco. 
      Electronic address: a.moumen@mascir.com.
LA  - eng
PT  - Journal Article
DEP - 20161228
PL  - Netherlands
TA  - Gene
JT  - Gene
JID - 7706761
RN  - 0 (RPL37A protein, human)
RN  - 0 (Ribosomal Proteins)
RN  - EC 2.7.10.1 (ERBB2 protein, human)
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Breast Neoplasms/*diagnosis/genetics/metabolism/pathology
MH  - Cell Line, Tumor
MH  - Female
MH  - Gene Expression
MH  - *Genes, Essential
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - Paraffin Embedding
MH  - Real-Time Polymerase Chain Reaction/*standards
MH  - Receptor, ErbB-2/*genetics/metabolism
MH  - Reference Standards
MH  - Ribosomal Proteins/*genetics/metabolism
MH  - Sensitivity and Specificity
MH  - Tissue Fixation
OTO - NOTNLM
OT  - Breast cancer
OT  - Endogenous control genes
OT  - HER2 expression
OT  - Molecular diagnosis
OT  - RT-qPCR
EDAT- 2017/01/01 06:00
MHDA- 2017/02/07 06:00
CRDT- 2017/01/01 06:00
PHST- 2016/06/16 00:00 [received]
PHST- 2016/12/08 00:00 [revised]
PHST- 2016/12/23 00:00 [accepted]
PHST- 2017/01/01 06:00 [pubmed]
PHST- 2017/02/07 06:00 [medline]
PHST- 2017/01/01 06:00 [entrez]
AID - S0378-1119(16)31027-7 [pii]
AID - 10.1016/j.gene.2016.12.027 [doi]
PST - ppublish
SO  - Gene. 2017 Mar 20;605:114-122. doi: 10.1016/j.gene.2016.12.027. Epub 2016 Dec 28.