Development of real-time quantitative reverse transcription-PCR for Her2 detection in peripheral blood from patients with breast cancer

Maria Savino, Maria Garrubba, Paola Parrella, Filomena Baorda, Massimiliano Copetti, Roberto Murgo, Leopoldo Zelante, Massimo Carella, Vanna Maria Valori, Stefano Angelo Santini

Research output: Contribution to journalArticle

Abstract

Background: Amplification of the Her2neu oncogene is a well known indicator of poor prognosis in breast cancer patients. The implementation into the clinical setting of therapeutical strategies directly targeting the Her2neu gene product, has create the need for the development of non-invasive analytical techniques in order to monitoring minimal residual disease and response to the treatment. Methods: To detect the expression of Her2neu mRNA in peripheral blood, we developed a specific real-time quantitative reverse transcription-PCR (QPCR) assay. The analyses were performed on blood samples obtained from 30 breast cancer patients positive for Her2neu overexpression by immunohistochemical analysis (IHC), 10 breast cancer patients negative for Her2neu overexpression, and 24 healthy controls. Results: Her2neu positive tumors showed a significant increase in mRNA transcripts as compared with both healthy controls (n = 24) and Her2neu negative patients (n = 10). After establishing a cut-off value, 18 out of the 30 Her2neu positive patient scored positive for Her2neu expression, whereas only 1 out of the 10 Her2neu negative patients was weakly positive. Conclusions: Her2neu QPCR is suitable method for Her2neu overexpression detection in peripheral blood from clinical samples of breast cancer patients. QPCR could be used to identify breast cancer patients with poor prognosis and for monitoring response to the therapy.

Original languageEnglish
Pages (from-to)52-56
Number of pages5
JournalClinica Chimica Acta
Volume384
Issue number1-2
DOIs
Publication statusPublished - Sep 2007

Fingerprint

Transcription
Reverse Transcription
Blood
Breast Neoplasms
Polymerase Chain Reaction
Messenger RNA
Monitoring
Amplification
Tumors
Assays
Genes
Gene Targeting
Residual Neoplasm
Oncogenes
Therapeutics

Keywords

  • Breast cancer
  • HER2
  • IHC
  • Quantitative PCR
  • Tumoral markers

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Development of real-time quantitative reverse transcription-PCR for Her2 detection in peripheral blood from patients with breast cancer. / Savino, Maria; Garrubba, Maria; Parrella, Paola; Baorda, Filomena; Copetti, Massimiliano; Murgo, Roberto; Zelante, Leopoldo; Carella, Massimo; Valori, Vanna Maria; Santini, Stefano Angelo.

In: Clinica Chimica Acta, Vol. 384, No. 1-2, 09.2007, p. 52-56.

Research output: Contribution to journalArticle

Savino, Maria ; Garrubba, Maria ; Parrella, Paola ; Baorda, Filomena ; Copetti, Massimiliano ; Murgo, Roberto ; Zelante, Leopoldo ; Carella, Massimo ; Valori, Vanna Maria ; Santini, Stefano Angelo. / Development of real-time quantitative reverse transcription-PCR for Her2 detection in peripheral blood from patients with breast cancer. In: Clinica Chimica Acta. 2007 ; Vol. 384, No. 1-2. pp. 52-56.
@article{0b4a667c27534ad486f8a347cb5c2e05,
title = "Development of real-time quantitative reverse transcription-PCR for Her2 detection in peripheral blood from patients with breast cancer",
abstract = "Background: Amplification of the Her2neu oncogene is a well known indicator of poor prognosis in breast cancer patients. The implementation into the clinical setting of therapeutical strategies directly targeting the Her2neu gene product, has create the need for the development of non-invasive analytical techniques in order to monitoring minimal residual disease and response to the treatment. Methods: To detect the expression of Her2neu mRNA in peripheral blood, we developed a specific real-time quantitative reverse transcription-PCR (QPCR) assay. The analyses were performed on blood samples obtained from 30 breast cancer patients positive for Her2neu overexpression by immunohistochemical analysis (IHC), 10 breast cancer patients negative for Her2neu overexpression, and 24 healthy controls. Results: Her2neu positive tumors showed a significant increase in mRNA transcripts as compared with both healthy controls (n = 24) and Her2neu negative patients (n = 10). After establishing a cut-off value, 18 out of the 30 Her2neu positive patient scored positive for Her2neu expression, whereas only 1 out of the 10 Her2neu negative patients was weakly positive. Conclusions: Her2neu QPCR is suitable method for Her2neu overexpression detection in peripheral blood from clinical samples of breast cancer patients. QPCR could be used to identify breast cancer patients with poor prognosis and for monitoring response to the therapy.",
keywords = "Breast cancer, HER2, IHC, Quantitative PCR, Tumoral markers",
author = "Maria Savino and Maria Garrubba and Paola Parrella and Filomena Baorda and Massimiliano Copetti and Roberto Murgo and Leopoldo Zelante and Massimo Carella and Valori, {Vanna Maria} and Santini, {Stefano Angelo}",
year = "2007",
month = "9",
doi = "10.1016/j.cca.2007.05.023",
language = "English",
volume = "384",
pages = "52--56",
journal = "Clinica Chimica Acta",
issn = "0009-8981",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Development of real-time quantitative reverse transcription-PCR for Her2 detection in peripheral blood from patients with breast cancer

AU - Savino, Maria

AU - Garrubba, Maria

AU - Parrella, Paola

AU - Baorda, Filomena

AU - Copetti, Massimiliano

AU - Murgo, Roberto

AU - Zelante, Leopoldo

AU - Carella, Massimo

AU - Valori, Vanna Maria

AU - Santini, Stefano Angelo

PY - 2007/9

Y1 - 2007/9

N2 - Background: Amplification of the Her2neu oncogene is a well known indicator of poor prognosis in breast cancer patients. The implementation into the clinical setting of therapeutical strategies directly targeting the Her2neu gene product, has create the need for the development of non-invasive analytical techniques in order to monitoring minimal residual disease and response to the treatment. Methods: To detect the expression of Her2neu mRNA in peripheral blood, we developed a specific real-time quantitative reverse transcription-PCR (QPCR) assay. The analyses were performed on blood samples obtained from 30 breast cancer patients positive for Her2neu overexpression by immunohistochemical analysis (IHC), 10 breast cancer patients negative for Her2neu overexpression, and 24 healthy controls. Results: Her2neu positive tumors showed a significant increase in mRNA transcripts as compared with both healthy controls (n = 24) and Her2neu negative patients (n = 10). After establishing a cut-off value, 18 out of the 30 Her2neu positive patient scored positive for Her2neu expression, whereas only 1 out of the 10 Her2neu negative patients was weakly positive. Conclusions: Her2neu QPCR is suitable method for Her2neu overexpression detection in peripheral blood from clinical samples of breast cancer patients. QPCR could be used to identify breast cancer patients with poor prognosis and for monitoring response to the therapy.

AB - Background: Amplification of the Her2neu oncogene is a well known indicator of poor prognosis in breast cancer patients. The implementation into the clinical setting of therapeutical strategies directly targeting the Her2neu gene product, has create the need for the development of non-invasive analytical techniques in order to monitoring minimal residual disease and response to the treatment. Methods: To detect the expression of Her2neu mRNA in peripheral blood, we developed a specific real-time quantitative reverse transcription-PCR (QPCR) assay. The analyses were performed on blood samples obtained from 30 breast cancer patients positive for Her2neu overexpression by immunohistochemical analysis (IHC), 10 breast cancer patients negative for Her2neu overexpression, and 24 healthy controls. Results: Her2neu positive tumors showed a significant increase in mRNA transcripts as compared with both healthy controls (n = 24) and Her2neu negative patients (n = 10). After establishing a cut-off value, 18 out of the 30 Her2neu positive patient scored positive for Her2neu expression, whereas only 1 out of the 10 Her2neu negative patients was weakly positive. Conclusions: Her2neu QPCR is suitable method for Her2neu overexpression detection in peripheral blood from clinical samples of breast cancer patients. QPCR could be used to identify breast cancer patients with poor prognosis and for monitoring response to the therapy.

KW - Breast cancer

KW - HER2

KW - IHC

KW - Quantitative PCR

KW - Tumoral markers

UR - http://www.scopus.com/inward/record.url?scp=34547850290&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34547850290&partnerID=8YFLogxK

U2 - 10.1016/j.cca.2007.05.023

DO - 10.1016/j.cca.2007.05.023

M3 - Article

C2 - 17599819

AN - SCOPUS:34547850290

VL - 384

SP - 52

EP - 56

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

SN - 0009-8981

IS - 1-2

ER -