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Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468
BACKGROUND: Increased transcription of oncogenes like the epidermal growth factor receptor (EGFR) is frequently caused by amplification of the whole gene or at least of regulatory sequences. Aim of this study was to pinpoint mechanistic parameters occurring during egfr copy number gains leading to a...
Autores principales: | , , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2841141/ https://www.ncbi.nlm.nih.gov/pubmed/20199686 http://dx.doi.org/10.1186/1471-2407-10-78 |
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author | Agelopoulos, Konstantin Greve, Burkhard Schmidt, Hartmut Pospisil, Heike Kurtz, Stefan Bartkowiak, Kai Andreas, Antje Wieczorek, Marek Korsching, Eberhard Buerger, Horst Brandt, Burkhard |
author_facet | Agelopoulos, Konstantin Greve, Burkhard Schmidt, Hartmut Pospisil, Heike Kurtz, Stefan Bartkowiak, Kai Andreas, Antje Wieczorek, Marek Korsching, Eberhard Buerger, Horst Brandt, Burkhard |
author_sort | Agelopoulos, Konstantin |
collection | PubMed |
description | BACKGROUND: Increased transcription of oncogenes like the epidermal growth factor receptor (EGFR) is frequently caused by amplification of the whole gene or at least of regulatory sequences. Aim of this study was to pinpoint mechanistic parameters occurring during egfr copy number gains leading to a stable EGFR overexpression and high sensitivity to extracellular signalling. A deeper understanding of those marker events might improve early diagnosis of cancer in suspect lesions, early detection of cancer progression and the prediction of egfr targeted therapies. METHODS: The basal-like/stemness type breast cancer cell line subpopulation MDA-MB-468 CD44(high)/CD24(-/low), carrying high egfr amplifications, was chosen as a model system in this study. Subclones of the heterogeneous cell line expressing low and high EGF receptor densities were isolated by cell sorting. Genomic profiling was carried out for these by means of SNP array profiling, qPCR and FISH. Cell cycle analysis was performed using the BrdU quenching technique. RESULTS: Low and high EGFR expressing MDA-MB-468 CD44(+)/CD24(-/low )subpopulations separated by cell sorting showed intermediate and high copy numbers of egfr, respectively. However, during cell culture an increase solely for egfr gene copy numbers in the intermediate subpopulation occurred. This shift was based on the formation of new cells which regained egfr gene copies. By two parametric cell cycle analysis clonal effects mediated through growth advantage of cells bearing higher egfr gene copy numbers could most likely be excluded for being the driving force. Subsequently, the detection of a fragile site distal to the egfr gene, sustaining uncapped telomere-less chromosomal ends, the ladder-like structure of the intrachromosomal egfr amplification and a broader range of egfr copy numbers support the assumption that dynamic chromosomal rearrangements, like breakage-fusion-bridge-cycles other than proliferation drive the gain of egfr copies. CONCLUSION: Progressive genome modulation in the CD44(+)/CD24(-/low )subpopulation of the breast cancer cell line MDA-MB-468 leads to different coexisting subclones. In isolated low-copy cells asymmetric chromosomal segregation leads to new cells with regained solely egfr gene copies. Furthermore, egfr regain resulted in enhanced signal transduction of the MAP-kinase and PI3-kinase pathway. We show here for the first time a dynamic copy number regain in basal-like/stemness cell type breast cancer subpopulations which might explain genetic heterogeneity. Moreover, this process might also be involved in adaptive growth factor receptor intracellular signaling which support survival and migration during cancer development and progression. |
format | Text |
id | pubmed-2841141 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-28411412010-03-18 Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 Agelopoulos, Konstantin Greve, Burkhard Schmidt, Hartmut Pospisil, Heike Kurtz, Stefan Bartkowiak, Kai Andreas, Antje Wieczorek, Marek Korsching, Eberhard Buerger, Horst Brandt, Burkhard BMC Cancer Research Article BACKGROUND: Increased transcription of oncogenes like the epidermal growth factor receptor (EGFR) is frequently caused by amplification of the whole gene or at least of regulatory sequences. Aim of this study was to pinpoint mechanistic parameters occurring during egfr copy number gains leading to a stable EGFR overexpression and high sensitivity to extracellular signalling. A deeper understanding of those marker events might improve early diagnosis of cancer in suspect lesions, early detection of cancer progression and the prediction of egfr targeted therapies. METHODS: The basal-like/stemness type breast cancer cell line subpopulation MDA-MB-468 CD44(high)/CD24(-/low), carrying high egfr amplifications, was chosen as a model system in this study. Subclones of the heterogeneous cell line expressing low and high EGF receptor densities were isolated by cell sorting. Genomic profiling was carried out for these by means of SNP array profiling, qPCR and FISH. Cell cycle analysis was performed using the BrdU quenching technique. RESULTS: Low and high EGFR expressing MDA-MB-468 CD44(+)/CD24(-/low )subpopulations separated by cell sorting showed intermediate and high copy numbers of egfr, respectively. However, during cell culture an increase solely for egfr gene copy numbers in the intermediate subpopulation occurred. This shift was based on the formation of new cells which regained egfr gene copies. By two parametric cell cycle analysis clonal effects mediated through growth advantage of cells bearing higher egfr gene copy numbers could most likely be excluded for being the driving force. Subsequently, the detection of a fragile site distal to the egfr gene, sustaining uncapped telomere-less chromosomal ends, the ladder-like structure of the intrachromosomal egfr amplification and a broader range of egfr copy numbers support the assumption that dynamic chromosomal rearrangements, like breakage-fusion-bridge-cycles other than proliferation drive the gain of egfr copies. CONCLUSION: Progressive genome modulation in the CD44(+)/CD24(-/low )subpopulation of the breast cancer cell line MDA-MB-468 leads to different coexisting subclones. In isolated low-copy cells asymmetric chromosomal segregation leads to new cells with regained solely egfr gene copies. Furthermore, egfr regain resulted in enhanced signal transduction of the MAP-kinase and PI3-kinase pathway. We show here for the first time a dynamic copy number regain in basal-like/stemness cell type breast cancer subpopulations which might explain genetic heterogeneity. Moreover, this process might also be involved in adaptive growth factor receptor intracellular signaling which support survival and migration during cancer development and progression. BioMed Central 2010-03-03 /pmc/articles/PMC2841141/ /pubmed/20199686 http://dx.doi.org/10.1186/1471-2407-10-78 Text en Copyright ©2010 Agelopoulos et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Agelopoulos, Konstantin Greve, Burkhard Schmidt, Hartmut Pospisil, Heike Kurtz, Stefan Bartkowiak, Kai Andreas, Antje Wieczorek, Marek Korsching, Eberhard Buerger, Horst Brandt, Burkhard Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 |
title | Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 |
title_full | Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 |
title_fullStr | Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 |
title_full_unstemmed | Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 |
title_short | Selective regain of egfr gene copies in CD44(+)/CD24(-/low )breast cancer cellular model MDA-MB-468 |
title_sort | selective regain of egfr gene copies in cd44(+)/cd24(-/low )breast cancer cellular model mda-mb-468 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2841141/ https://www.ncbi.nlm.nih.gov/pubmed/20199686 http://dx.doi.org/10.1186/1471-2407-10-78 |
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