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Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells

Based on molecular features, breast cancers are grouped into intrinsic subtypes that have different prognoses and therapeutic response profiles. With increasing age, breast cancer incidence increases, with hormone receptor-positive and other luminal-like subtype tumors comprising a majority of cases...

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Autores principales: Lee, Jonathan K., Garbe, James C., Vrba, Lukas, Miyano, Masaru, Futscher, Bernard W., Stampfer, Martha R., LaBarge, Mark A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4356162/
https://www.ncbi.nlm.nih.gov/pubmed/25815289
http://dx.doi.org/10.3389/fcell.2015.00013
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author Lee, Jonathan K.
Garbe, James C.
Vrba, Lukas
Miyano, Masaru
Futscher, Bernard W.
Stampfer, Martha R.
LaBarge, Mark A.
author_facet Lee, Jonathan K.
Garbe, James C.
Vrba, Lukas
Miyano, Masaru
Futscher, Bernard W.
Stampfer, Martha R.
LaBarge, Mark A.
author_sort Lee, Jonathan K.
collection PubMed
description Based on molecular features, breast cancers are grouped into intrinsic subtypes that have different prognoses and therapeutic response profiles. With increasing age, breast cancer incidence increases, with hormone receptor-positive and other luminal-like subtype tumors comprising a majority of cases. It is not known at what stage of tumor progression subtype specification occurs, nor how the process of aging affects the intrinsic subtype. We examined subtype markers in immortalized human mammary epithelial cell lines established following exposure of primary cultured cell strains to a two-step immortalization protocol that targets the two main barriers to immortality: stasis (stress-associated senescence) and replicative senescence. Cell lines derived from epithelial cells obtained from non-tumorous pre- and post-menopausal breast surgery tissues were compared. Additionally, comparisons were made between lines generated using two different genetic interventions to bypass stasis: transduction of either an shRNA that down-regulated p16(INK4A), or overexpressed constitutive active cyclin D1/CDK2. In all cases, the replicative senescence barrier was bypassed by transduction of c-Myc. Cells from all resulting immortal lines exhibited normal karyotypes. Immunofluorescence, flow cytometry, and gene expression analyses of lineage-specific markers were used to categorize the intrinsic subtypes of the immortalized lines. Bypassing stasis with p16 shRNA in young strains generated cell lines that were invariably basal-like, but the lines examined from older strains exhibited some luminal features such as keratin 19 and estrogen receptor expression. Overexpression of cyclin D1/CDK2 resulted in keratin 19 positive, luminal-like cell lines from both young and old strains, and the lines examined from older strains exhibited estrogen receptor expression. Thus age and the method of bypassing stasis independently influence the subtype of immortalized human mammary epithelial cells.
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spelling pubmed-43561622015-03-26 Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells Lee, Jonathan K. Garbe, James C. Vrba, Lukas Miyano, Masaru Futscher, Bernard W. Stampfer, Martha R. LaBarge, Mark A. Front Cell Dev Biol Cell and Developmental Biology Based on molecular features, breast cancers are grouped into intrinsic subtypes that have different prognoses and therapeutic response profiles. With increasing age, breast cancer incidence increases, with hormone receptor-positive and other luminal-like subtype tumors comprising a majority of cases. It is not known at what stage of tumor progression subtype specification occurs, nor how the process of aging affects the intrinsic subtype. We examined subtype markers in immortalized human mammary epithelial cell lines established following exposure of primary cultured cell strains to a two-step immortalization protocol that targets the two main barriers to immortality: stasis (stress-associated senescence) and replicative senescence. Cell lines derived from epithelial cells obtained from non-tumorous pre- and post-menopausal breast surgery tissues were compared. Additionally, comparisons were made between lines generated using two different genetic interventions to bypass stasis: transduction of either an shRNA that down-regulated p16(INK4A), or overexpressed constitutive active cyclin D1/CDK2. In all cases, the replicative senescence barrier was bypassed by transduction of c-Myc. Cells from all resulting immortal lines exhibited normal karyotypes. Immunofluorescence, flow cytometry, and gene expression analyses of lineage-specific markers were used to categorize the intrinsic subtypes of the immortalized lines. Bypassing stasis with p16 shRNA in young strains generated cell lines that were invariably basal-like, but the lines examined from older strains exhibited some luminal features such as keratin 19 and estrogen receptor expression. Overexpression of cyclin D1/CDK2 resulted in keratin 19 positive, luminal-like cell lines from both young and old strains, and the lines examined from older strains exhibited estrogen receptor expression. Thus age and the method of bypassing stasis independently influence the subtype of immortalized human mammary epithelial cells. Frontiers Media S.A. 2015-03-11 /pmc/articles/PMC4356162/ /pubmed/25815289 http://dx.doi.org/10.3389/fcell.2015.00013 Text en Copyright © 2015 Lee, Garbe, Vrba, Miyano, Futscher, Stampfer and LaBarge. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Lee, Jonathan K.
Garbe, James C.
Vrba, Lukas
Miyano, Masaru
Futscher, Bernard W.
Stampfer, Martha R.
LaBarge, Mark A.
Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
title Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
title_full Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
title_fullStr Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
title_full_unstemmed Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
title_short Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
title_sort age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4356162/
https://www.ncbi.nlm.nih.gov/pubmed/25815289
http://dx.doi.org/10.3389/fcell.2015.00013
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