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Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies

BACKGROUND: Prolactin is a polypeptide hormone responsible for proliferation and differentiation of the mammary gland. More recently, prolactin's role in mammary carcinogenesis has been studied with greater interest. Studies from our laboratory and from others have demonstrated that three speci...

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Autores principales: Ginsburg, Erika, Alexander, Stefanie, Lieber, Sarah, Tarplin, Sarah, Jenkins, Luwanda, Pang, Linda, Heger, Christopher D, Goldsmith, Paul, Vonderhaar, Barbara K
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3009681/
https://www.ncbi.nlm.nih.gov/pubmed/21144038
http://dx.doi.org/10.1186/1471-2407-10-678
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author Ginsburg, Erika
Alexander, Stefanie
Lieber, Sarah
Tarplin, Sarah
Jenkins, Luwanda
Pang, Linda
Heger, Christopher D
Goldsmith, Paul
Vonderhaar, Barbara K
author_facet Ginsburg, Erika
Alexander, Stefanie
Lieber, Sarah
Tarplin, Sarah
Jenkins, Luwanda
Pang, Linda
Heger, Christopher D
Goldsmith, Paul
Vonderhaar, Barbara K
author_sort Ginsburg, Erika
collection PubMed
description BACKGROUND: Prolactin is a polypeptide hormone responsible for proliferation and differentiation of the mammary gland. More recently, prolactin's role in mammary carcinogenesis has been studied with greater interest. Studies from our laboratory and from others have demonstrated that three specific isoforms of the prolactin receptor (PRLR) are expressed in both normal and cancerous breast cells and tissues. Until now, reliable isoform specific antibodies have been lacking. We have prepared and characterized polyclonal antibodies against each of the human PRLR isoforms that can effectively be used to characterize human breast cancers. METHODS: Rabbits were immunized with synthetic peptides of isoform unique regions and immune sera affinity purified prior to validation by Western blot and immunohistochemical analyses. Sections of ductal and lobular carcinomas were stained with each affinity purified isoform specific antibody to determine expression patterns in breast cancer subclasses. RESULTS: We show that the rabbit antibodies have high titer and could specifically recognize each isoform of PRLR. Differences in PRLR isoform expression levels were observed and quantified using histosections from xenografts of established human breast cancer cells lines, and ductal and lobular carcinoma human biopsy specimens. In addition, these results were verified by real-time PCR with isoform specific primers. While nearly all tumors contained LF and SF1b, the majority (76%) of ductal carcinoma biopsies expressed SF1a while the majority of lobular carcinomas lacked SF1a staining (72%) and 27% had only low levels of expression. CONCLUSIONS: Differences in the receptor isoform expression profiles may be critical to understanding the role of PRL in mammary tumorigenesis. Since these antibodies are specifically directed against each PRLR isoform, they are valuable tools for the evaluation of breast cancer PRLR content and have potential clinical importance in treatment of this disease by providing new reagents to study the protein expression of the human PRLR.
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spelling pubmed-30096812010-12-24 Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies Ginsburg, Erika Alexander, Stefanie Lieber, Sarah Tarplin, Sarah Jenkins, Luwanda Pang, Linda Heger, Christopher D Goldsmith, Paul Vonderhaar, Barbara K BMC Cancer Research Article BACKGROUND: Prolactin is a polypeptide hormone responsible for proliferation and differentiation of the mammary gland. More recently, prolactin's role in mammary carcinogenesis has been studied with greater interest. Studies from our laboratory and from others have demonstrated that three specific isoforms of the prolactin receptor (PRLR) are expressed in both normal and cancerous breast cells and tissues. Until now, reliable isoform specific antibodies have been lacking. We have prepared and characterized polyclonal antibodies against each of the human PRLR isoforms that can effectively be used to characterize human breast cancers. METHODS: Rabbits were immunized with synthetic peptides of isoform unique regions and immune sera affinity purified prior to validation by Western blot and immunohistochemical analyses. Sections of ductal and lobular carcinomas were stained with each affinity purified isoform specific antibody to determine expression patterns in breast cancer subclasses. RESULTS: We show that the rabbit antibodies have high titer and could specifically recognize each isoform of PRLR. Differences in PRLR isoform expression levels were observed and quantified using histosections from xenografts of established human breast cancer cells lines, and ductal and lobular carcinoma human biopsy specimens. In addition, these results were verified by real-time PCR with isoform specific primers. While nearly all tumors contained LF and SF1b, the majority (76%) of ductal carcinoma biopsies expressed SF1a while the majority of lobular carcinomas lacked SF1a staining (72%) and 27% had only low levels of expression. CONCLUSIONS: Differences in the receptor isoform expression profiles may be critical to understanding the role of PRL in mammary tumorigenesis. Since these antibodies are specifically directed against each PRLR isoform, they are valuable tools for the evaluation of breast cancer PRLR content and have potential clinical importance in treatment of this disease by providing new reagents to study the protein expression of the human PRLR. BioMed Central 2010-12-13 /pmc/articles/PMC3009681/ /pubmed/21144038 http://dx.doi.org/10.1186/1471-2407-10-678 Text en Copyright ©2010 Ginsburg et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (<url>http://creativecommons.org/licenses/by/2.0</url>), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ginsburg, Erika
Alexander, Stefanie
Lieber, Sarah
Tarplin, Sarah
Jenkins, Luwanda
Pang, Linda
Heger, Christopher D
Goldsmith, Paul
Vonderhaar, Barbara K
Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
title Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
title_full Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
title_fullStr Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
title_full_unstemmed Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
title_short Characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
title_sort characterization of ductal and lobular breast carcinomas using novel prolactin receptor isoform specific antibodies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3009681/
https://www.ncbi.nlm.nih.gov/pubmed/21144038
http://dx.doi.org/10.1186/1471-2407-10-678
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