title: Quantitative Real Time PCR assessment of hormonal receptors and HER2 status on fine-needle aspiration pre-operatory specimens from a prospectively accrued cohort of women with suspect breast malignant lesions.
abstract: OBJECTIVE
  Reliable assessment of estrogen, progesterone (ER and PR), and HER2 receptor status are essential in breast cancer (BC) treatment. Immunohistochemical methods are limited by intra- and inter-laboratory variability. Furthermore, current methods are not the ideal approach for reproducing the biological continuum of ER, PR, and HER2 receptor levels, due to their intrinsic, semi-quantitative nature, relying in part on subjective interpretation.
  
  
  METHODS
  In the present study, we tested a molecular approach to define ER, PR, and HER2 status in fine-needle-aspirate (FNA) samples from patients with early BC. We performed flow cytometry analysis on 88 FNA specimens from suspect BC patients to determine cellularity. We used quantitative Real Time PCR (QRT-PCR) to assess ER, PR, HER2 status, and qPCR for HER2 gene copy number (GCN).
  
  
  RESULTS
  ER and PR mRNA levels showed a highly significant correlation with IHC data on surgical samples. qPCR showed greater accuracy than IHC in defining HER2 status. QRT-PCR defined better than IHC the continuous spectrum of the expression of the assessed receptors. Moreover, PCR analysis demonstrated a strict correlation between HER2 status and higher levels of its transcript, correctly stratifying HER2+ and HER2- patients. Finally, there was a strongly significant agreement between HER2 GCN assessed on FNA specimens by qPCR and FISH data obtained on pathological tissue specimens.
  
  
  CONCLUSIONS
  The present results support a comprehensive approach to determine ER, PR, and HER2 status by PCR (QRT-PCR and qPCR) in FNA specimens, with high relevance for therapeutic strategies like neoadjuvant treatment.
authors.affiliation: Department of Internal Medicine, Istituto di Ricerca a Carattere Clinico e Scientifico (IRCCS), Azienda Ospedaliera Universitaria (AOU) San Martino Istituto Nazionale Tumori (IST), Genoa, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Garuti
authors.affiliation: Department of Internal Medicine, Istituto di Ricerca a Carattere Clinico e Scientifico (IRCCS), Azienda Ospedaliera Universitaria (AOU) San Martino Istituto Nazionale Tumori (IST), Genoa, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Rocco
authors.affiliation: Department of Internal Medicine, Istituto di Ricerca a Carattere Clinico e Scientifico (IRCCS), Azienda Ospedaliera Universitaria (AOU) San Martino Istituto Nazionale Tumori (IST), Genoa, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Cirmena
authors.affiliation: Division of Pathology, Busto Arsizio Hospital, Busto Arsizio, Varese, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Chiaramondia
authors.affiliation: Department of Surgical Sciences and Integrated Diagnostics, University of Genoa and Anatomic Pathology Service, IRCCS AOU San Martino IST, Genoa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Baccini
authors.affiliation: Department of Radiology, IRCCS AOU San Martino IST, Genoa, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Calabrese
authors.affiliation: Department of Internal Medicine, Istituto di Ricerca a Carattere Clinico e Scientifico (IRCCS), Azienda Ospedaliera Universitaria (AOU) San Martino Istituto Nazionale Tumori (IST), Genoa, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Palermo
authors.affiliation: Breast Surgery Unit, Department of Surgery, University of Genoa and IRCCS AOU San Martino IST, Genoa, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Friedman
authors.affiliation: Department of Internal Medicine, Istituto di Ricerca a Carattere Clinico e Scientifico (IRCCS), Azienda Ospedaliera Universitaria (AOU) San Martino Istituto Nazionale Tumori (IST), Genoa, Italy. Electronic address: gabriele.zoppoli@unige.it.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Zoppoli
authors.affiliation: Department of Internal Medicine, Istituto di Ricerca a Carattere Clinico e Scientifico (IRCCS), Azienda Ospedaliera Universitaria (AOU) San Martino Istituto Nazionale Tumori (IST), Genoa, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ballestrero
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D044963:Biopsy, Fine-Needle; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015331:Cohort Studies; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D011446:Prospective Studies; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000235:genetics / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone
keywords: Breast cancer; Fine needle aspiration; HER2; Hormonal receptors; PCR