Correlation of GREB1 mRNA with protein expression in breast cancer: Validation of a novel GREB1 monoclonal antibody

H. J. Hnatyszyn, M. Liu, A. Hilger, L. Herbert, Carmen Gomez-Fernandez, Merce Jorda, D. Thomas, J. M. Rae, Dorraya El-Ashry, Marc E Lippman

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Studies of gene regulated by estrogen in breast cancer 1 (GREB1) have focused on mRNA levels with limited evidence about GREB1 protein expression in normal and breast cancer cells. A monoclonal antibody that recognizes GREB1 protein in breast tissues could be applied to correlate protein expression with established mRNA expression data. A hybridoma expressing a murine monoclonal antibody targeting a 119 amino acid peptide specific to human GREB1 was generated. The novel monoclonal GREB1 antibody (GREB1ab) was validated for use in Western blotting as well as immunohistochemical (IHC) applications. GREB1ab detects a 216 kDa protein corresponding to GREB1 in estrogen receptor alpha (ERaα+) breast cancer cells as well as ERα- breast cancer cells transduced with a GREB1 expression vector. GREB1ab specificity was verified using an ERa antagonist to prevent GREB1 induction as well as a silencing siRNA targeting GREB1 mRNA. GREB1ab was further validated for detection of GREB1 by IHC in breast cancer cell lines and breast tissue microarrays (TMA). ERaα+cell lines were observed to express GREB1 while ERα- cell lines did not express detectable levels of the protein. Using breast cancer tissue whole sections, IHC with the GREB1ab identified protein expression in ERaα+ breast cancer tissue as well as normal breast tissue, with little GREB1 expression in ERα- breast cancer tissue. Furthermore, these data indicate that GREB1 mRNA expression correlates well with protein expression. The novel monoclonal GREB1ab is specific for GREB1 protein. This antibody will serve as a tool for investigations focused on the expression, distribution, and function of GREB1 in normal breast and breast cancer tissues.

Original languageEnglish
Pages (from-to)371-380
Number of pages10
JournalBreast Cancer Research and Treatment
Volume122
Issue number2
DOIs
StatePublished - Jul 1 2010

Fingerprint

Estrogens
Monoclonal Antibodies
Breast Neoplasms
Messenger RNA
Genes
Proteins
BRCA1 Protein
Breast
Antibodies
Cell Line
Antibody Specificity
Gene Targeting
Estrogen Receptor alpha
Hybridomas
Small Interfering RNA

Keywords

  • GREB1
  • Immunohistochemistry
  • Monoclonal antibody
  • mRNA-Protein correlation.

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Correlation of GREB1 mRNA with protein expression in breast cancer : Validation of a novel GREB1 monoclonal antibody. / Hnatyszyn, H. J.; Liu, M.; Hilger, A.; Herbert, L.; Gomez-Fernandez, Carmen; Jorda, Merce; Thomas, D.; Rae, J. M.; El-Ashry, Dorraya; Lippman, Marc E.

In: Breast Cancer Research and Treatment, Vol. 122, No. 2, 01.07.2010, p. 371-380.

Research output: Contribution to journalArticle

Hnatyszyn, H. J. ; Liu, M. ; Hilger, A. ; Herbert, L. ; Gomez-Fernandez, Carmen ; Jorda, Merce ; Thomas, D. ; Rae, J. M. ; El-Ashry, Dorraya ; Lippman, Marc E. / Correlation of GREB1 mRNA with protein expression in breast cancer : Validation of a novel GREB1 monoclonal antibody. In: Breast Cancer Research and Treatment. 2010 ; Vol. 122, No. 2. pp. 371-380.
@article{0f9a438241404bb29b32d4a93d8f5f3a,
title = "Correlation of GREB1 mRNA with protein expression in breast cancer: Validation of a novel GREB1 monoclonal antibody",
abstract = "Studies of gene regulated by estrogen in breast cancer 1 (GREB1) have focused on mRNA levels with limited evidence about GREB1 protein expression in normal and breast cancer cells. A monoclonal antibody that recognizes GREB1 protein in breast tissues could be applied to correlate protein expression with established mRNA expression data. A hybridoma expressing a murine monoclonal antibody targeting a 119 amino acid peptide specific to human GREB1 was generated. The novel monoclonal GREB1 antibody (GREB1ab) was validated for use in Western blotting as well as immunohistochemical (IHC) applications. GREB1ab detects a 216 kDa protein corresponding to GREB1 in estrogen receptor alpha (ERaα+) breast cancer cells as well as ERα- breast cancer cells transduced with a GREB1 expression vector. GREB1ab specificity was verified using an ERa antagonist to prevent GREB1 induction as well as a silencing siRNA targeting GREB1 mRNA. GREB1ab was further validated for detection of GREB1 by IHC in breast cancer cell lines and breast tissue microarrays (TMA). ERaα+cell lines were observed to express GREB1 while ERα- cell lines did not express detectable levels of the protein. Using breast cancer tissue whole sections, IHC with the GREB1ab identified protein expression in ERaα+ breast cancer tissue as well as normal breast tissue, with little GREB1 expression in ERα- breast cancer tissue. Furthermore, these data indicate that GREB1 mRNA expression correlates well with protein expression. The novel monoclonal GREB1ab is specific for GREB1 protein. This antibody will serve as a tool for investigations focused on the expression, distribution, and function of GREB1 in normal breast and breast cancer tissues.",
keywords = "GREB1, Immunohistochemistry, Monoclonal antibody, mRNA-Protein correlation.",
author = "Hnatyszyn, {H. J.} and M. Liu and A. Hilger and L. Herbert and Carmen Gomez-Fernandez and Merce Jorda and D. Thomas and Rae, {J. M.} and Dorraya El-Ashry and Lippman, {Marc E}",
year = "2010",
month = "7",
day = "1",
doi = "10.1007/s10549-009-0584-x",
language = "English",
volume = "122",
pages = "371--380",
journal = "Breast Cancer Research and Treatment",
issn = "0167-6806",
publisher = "Springer New York",
number = "2",

}

TY - JOUR

T1 - Correlation of GREB1 mRNA with protein expression in breast cancer

T2 - Validation of a novel GREB1 monoclonal antibody

AU - Hnatyszyn, H. J.

AU - Liu, M.

AU - Hilger, A.

AU - Herbert, L.

AU - Gomez-Fernandez, Carmen

AU - Jorda, Merce

AU - Thomas, D.

AU - Rae, J. M.

AU - El-Ashry, Dorraya

AU - Lippman, Marc E

PY - 2010/7/1

Y1 - 2010/7/1

N2 - Studies of gene regulated by estrogen in breast cancer 1 (GREB1) have focused on mRNA levels with limited evidence about GREB1 protein expression in normal and breast cancer cells. A monoclonal antibody that recognizes GREB1 protein in breast tissues could be applied to correlate protein expression with established mRNA expression data. A hybridoma expressing a murine monoclonal antibody targeting a 119 amino acid peptide specific to human GREB1 was generated. The novel monoclonal GREB1 antibody (GREB1ab) was validated for use in Western blotting as well as immunohistochemical (IHC) applications. GREB1ab detects a 216 kDa protein corresponding to GREB1 in estrogen receptor alpha (ERaα+) breast cancer cells as well as ERα- breast cancer cells transduced with a GREB1 expression vector. GREB1ab specificity was verified using an ERa antagonist to prevent GREB1 induction as well as a silencing siRNA targeting GREB1 mRNA. GREB1ab was further validated for detection of GREB1 by IHC in breast cancer cell lines and breast tissue microarrays (TMA). ERaα+cell lines were observed to express GREB1 while ERα- cell lines did not express detectable levels of the protein. Using breast cancer tissue whole sections, IHC with the GREB1ab identified protein expression in ERaα+ breast cancer tissue as well as normal breast tissue, with little GREB1 expression in ERα- breast cancer tissue. Furthermore, these data indicate that GREB1 mRNA expression correlates well with protein expression. The novel monoclonal GREB1ab is specific for GREB1 protein. This antibody will serve as a tool for investigations focused on the expression, distribution, and function of GREB1 in normal breast and breast cancer tissues.

AB - Studies of gene regulated by estrogen in breast cancer 1 (GREB1) have focused on mRNA levels with limited evidence about GREB1 protein expression in normal and breast cancer cells. A monoclonal antibody that recognizes GREB1 protein in breast tissues could be applied to correlate protein expression with established mRNA expression data. A hybridoma expressing a murine monoclonal antibody targeting a 119 amino acid peptide specific to human GREB1 was generated. The novel monoclonal GREB1 antibody (GREB1ab) was validated for use in Western blotting as well as immunohistochemical (IHC) applications. GREB1ab detects a 216 kDa protein corresponding to GREB1 in estrogen receptor alpha (ERaα+) breast cancer cells as well as ERα- breast cancer cells transduced with a GREB1 expression vector. GREB1ab specificity was verified using an ERa antagonist to prevent GREB1 induction as well as a silencing siRNA targeting GREB1 mRNA. GREB1ab was further validated for detection of GREB1 by IHC in breast cancer cell lines and breast tissue microarrays (TMA). ERaα+cell lines were observed to express GREB1 while ERα- cell lines did not express detectable levels of the protein. Using breast cancer tissue whole sections, IHC with the GREB1ab identified protein expression in ERaα+ breast cancer tissue as well as normal breast tissue, with little GREB1 expression in ERα- breast cancer tissue. Furthermore, these data indicate that GREB1 mRNA expression correlates well with protein expression. The novel monoclonal GREB1ab is specific for GREB1 protein. This antibody will serve as a tool for investigations focused on the expression, distribution, and function of GREB1 in normal breast and breast cancer tissues.

KW - GREB1

KW - Immunohistochemistry

KW - Monoclonal antibody

KW - mRNA-Protein correlation.

UR - http://www.scopus.com/inward/record.url?scp=77955655628&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955655628&partnerID=8YFLogxK

U2 - 10.1007/s10549-009-0584-x

DO - 10.1007/s10549-009-0584-x

M3 - Article

C2 - 19842031

AN - SCOPUS:77955655628

VL - 122

SP - 371

EP - 380

JO - Breast Cancer Research and Treatment

JF - Breast Cancer Research and Treatment

SN - 0167-6806

IS - 2

ER -