title: Evaluation of the prognostic significance of HER family mRNA expression in high-risk early breast cancer: a Hellenic Cooperative Oncology Group (HeCOG) validation study.
abstract: BACKGROUND
  The aim of the study was to evaluate the prognostic ability of the transcriptional profiling of the HER family genes in early breast cancer, as a validation analysis of another previously published HeCOG study.
  
  
  METHODS
  RNA was extracted from 663 formalin-fixed paraffin-embedded (FFPE) tumor tissue samples of high-risk early breast cancer patients enrolled in the randomized HE10/00 trial. Relative mRNA expression of all four HER family members was assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
  
  
  RESULTS
  In compliance with our previous study, the overall agreement between qRT-PCR and IHC/FISH for HER2 status determination was good (69%). Likewise, the overall concordance between qRT-PCR and IHC for EGFR status was high (81%). In line with our previously reported data, we demonstrated a positive association between HER2 and HER3 mRNA expression. Similarly, mRNA expression of HER3 and HER4 was positively associated with each other and negatively associated with EGFR. Regarding relationships with clinico-pathological parameters, our findings are also in agreement with our previous results. Generally, increased EGFR and HER2 mRNA expression was related to unfavorable, whereas high HER3 and HER4 mRNA expression was associated with favorable clinico-pathological parameters. In univariate analysis, no significant association between EGFR, HER2 and HER3 mRNA expression and overall survival (OS) or disease-free survival (DFS) was demonstrated. However, high EGFR protein expression was associated with significantly shorter OS (log-rank, p = 0.015). In compliance with our previously published data, increased HER4 mRNA expression had a significantly favorable prognostic value in terms of OS (p = 0.044) and DFS (p = 0.047). In multivariate analysis, among all HER receptors, only EGFR protein expression was found to affect OS (Wald's p = 0.028) and DFS (p = 0.015) independently. Concerning the combined expression of all four HER family receptors, the combination of high EGFR, high HER2, low HER3 and low HER4 mRNA expression was associated with a trend for shorter OS (log-rank, p = 0.065) and significantly worse DFS (p = 0.033), compared with all other co-expression profiles.
  
  
  CONCLUSIONS
  These data indicate that qRT-PCR may represent a valid alternative method for evaluating the expression of HER family members in FFPE breast carcinoma tissue samples.
  
  
  BACKGROUND
  Australian New Zealand Clinical Trials Registry ACTRN12609001036202.
authors.affiliation: Division of Oncology, Department of Medicine, University Hospital, University of Patras Medical School, 26504, Rio, Greece. angkoutr@otenet.gr.
authors.forename: Angelos
authors.initials: A
authors.lastname: Koutras
authors.affiliation: Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research, Aristotle University of Thessaloniki School of Medicine, University Campus, Bldg. 17B, 54 006, Thessaloniki, Greece. k_kalogeras@hecog.ondsl.gr.|Translational Research Section, Hellenic Cooperative Oncology Group, Data Office, 18 Hatzikonstanti Str., 115 24, Athens, Greece. k_kalogeras@hecog.ondsl.gr.
authors.forename: Konstantine T
authors.initials: KT
authors.lastname: Kalogeras
authors.affiliation: STRATIFYER Molecular Pathology GmbH, Werthmannstr. 1c, 50935, Cologne, Germany. ralph.wirtz@stratifyer.de.
authors.forename: Ralph M
authors.initials: RM
authors.lastname: Wirtz
authors.affiliation: Health Data Specialists Ltd, 22 Katehaki Str., 115 25, Athens, Greece. zoi.alexopoulou@heads.gr.
authors.forename: Zoi
authors.initials: Z
authors.lastname: Alexopoulou
authors.affiliation: Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research, Aristotle University of Thessaloniki School of Medicine, University Campus, Bldg. 17B, 54 006, Thessaloniki, Greece. mbobos@auth.gr.
authors.forename: Mattheos
authors.initials: M
authors.lastname: Bobos
authors.affiliation: Department of Clinical Therapeutics, "Alexandra" Hospital, University of Athens School of Medicine, 80 Vasilissis Sofias Av. and Lourou Str., 115 28, Athens, Greece. florazagouri@yahoo.co.uk.
authors.forename: Flora
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authors.lastname: Zagouri
authors.affiliation: STRATIFYER Molecular Pathology GmbH, Werthmannstr. 1c, 50935, Cologne, Germany. elke.veltrup@stratifyer.de.
authors.forename: Elke
authors.initials: E
authors.lastname: Veltrup
authors.affiliation: Department of Medical Oncology, "Papageorgiou" Hospital, Aristotle University of Thessaloniki School of Medicine, Ring Road, Nea Efkarpia, 56 429, Thessaloniki, Greece. nellitim@doctors.org.uk.
authors.forename: Eleni
authors.initials: E
authors.lastname: Timotheadou
authors.affiliation: First Department of Medicine, "Laiko" General Hospital, University of Athens School of Medicine, 17 Ag. Thoma Str., 115 27, Athens, Greece. hgogas@hol.gr.
authors.forename: Helen
authors.initials: H
authors.lastname: Gogas
authors.affiliation: Department of Medical Oncology, Ioannina University Hospital, Niarchou Av., 45 500, Ioannina, Greece. gpenther@otenet.gr.
authors.forename: George
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authors.lastname: Pentheroudakis
authors.affiliation: Department of Medical Oncology, IKA Hospital, 40 Mavromichali Str., 54248, Thessaloniki, Greece. n.pisanidis@gmail.com.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Pisanidis
authors.affiliation: Pathology Department, "Evangelismos" Hospital, 45 Ipsilantou Str., 106 76, Athens, Greece. cmagkou@yahoo.com.
authors.forename: Christina
authors.initials: C
authors.lastname: Magkou
authors.affiliation: Second Department of Medical Oncology, "Metropolitan" Hospital, 9 Ethnarchou Makariou Str. and El. Venizelou Str., 185 47, Piraeus, Greece. c_christodoulou@yahoo.gr.
authors.forename: Christos
authors.initials: C
authors.lastname: Christodoulou
authors.affiliation: First Department of Medical Oncology, "Metropolitan" Hospital, 9 Ethnarchou Makariou Str. and El. Venizelou Str., 185 47, Piraeus, Greece. dimmp@otenet.gr.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Bafaloukos
authors.affiliation: Oncology Unit, "Hippokration" Hospital, 108 Vas. Sofias Av., 115 27, Athens, Greece. p.papakostas@gmail.com.
authors.forename: Pavlos
authors.initials: P
authors.lastname: Papakostas
authors.affiliation: Second Department of Medical Oncology, "Agii Anargiri" Cancer Hospital, Noufaron Str. and Timiou Stavrou Str., 145 64, Athens, Greece. garavantinos@yahoo.gr.
authors.forename: Gerasimos
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authors.lastname: Aravantinos
authors.affiliation: Oncology Section, Second Department of Internal Medicine, "Hippokration" Hospital, 114 Vas. Sofias Av., 115 27, Athens, Greece. pectasid@otenet.gr.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Pectasides
authors.affiliation: Division of Oncology, Department of Medicine, University Hospital, University of Patras Medical School, 26504, Rio, Greece. kalofonos@upatras.gr.
authors.forename: Haralambos P
authors.initials: HP
authors.lastname: Kalofonos
authors.affiliation: Laboratory of Molecular Oncology, Hellenic Foundation for Cancer Research, Aristotle University of Thessaloniki School of Medicine, University Campus, Bldg. 17B, 54 006, Thessaloniki, Greece. fountzil@auth.gr.
authors.forename: George
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authors.lastname: Fountzilas
mesh_terms: D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000235:genetics*; D018572:Disease-Free Survival; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006115:Greece; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D020893:Receptor, ErbB-3 / Q000235:genetics / Q000378:metabolism; D066247:Receptor, ErbB-4 / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012307:Risk Factors; D055815:Young Adult
chemical_list: D066246:ErbB Receptors; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D020893:Receptor, ErbB-3; D066247:Receptor, ErbB-4