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

Anna Garuti, Ilaria Rocco, Gabriella Cirmena, Maurizio Chiaramondia, Paola Baccini, Massimo Calabrese, Claudia Palermo, Daniele Friedman, Gabriele Zoppoli, Alberto Ballestrero

Research output: Contribution to journalArticle

Abstract

Objectives 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.

Original languageEnglish
Pages (from-to)389-396
Number of pages8
JournalGynecologic Oncology
Volume132
Issue number2
DOIs
Publication statusPublished - Feb 2014

Fingerprint

Fine Needle Biopsy
Needles
Real-Time Polymerase Chain Reaction
Breast
erbB-2 Genes
Gene Dosage
Breast Neoplasms
Polymerase Chain Reaction
Neoadjuvant Therapy
Progesterone
Flow Cytometry
Estrogens
Messenger RNA
Therapeutics

Keywords

  • Breast cancer
  • Fine needle aspiration
  • HER2
  • Hormonal receptors
  • PCR

ASJC Scopus subject areas

  • Obstetrics and Gynaecology
  • Oncology

Cite this

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. / Garuti, Anna; Rocco, Ilaria; Cirmena, Gabriella; Chiaramondia, Maurizio; Baccini, Paola; Calabrese, Massimo; Palermo, Claudia; Friedman, Daniele; Zoppoli, Gabriele; Ballestrero, Alberto.

In: Gynecologic Oncology, Vol. 132, No. 2, 02.2014, p. 389-396.

Research output: Contribution to journalArticle

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abstract = "Objectives 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.",
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AU - Cirmena, Gabriella

AU - Chiaramondia, Maurizio

AU - Baccini, Paola

AU - Calabrese, Massimo

AU - Palermo, Claudia

AU - Friedman, Daniele

AU - Zoppoli, Gabriele

AU - Ballestrero, Alberto

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