Genetic and epigenetic analysis of putative breast cancer stem cell models

Marija Balic, Daniela Schwarzenbacher, Stefanie Stanzer, Ellen Heitzer, Martina Auer, Jochen B. Geigl, Richard J Cote, Ram Datar, Nadia Dandachi

Research output: Contribution to journalArticle

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Abstract

Background: Cancer stem cell model hypothesizes existence of a small proportion of tumor cells capable of sustaining tumor formation, self-renewal and differentiation. In breast cancer, these cells were found to be associated with CD44+CD24-low and ALDH+ phenotype. Our study was performed to evaluate the suitability of current approaches for breast cancer stem cell analyses to evaluate heterogeneity of breast cancer cells through their extensive genetic and epigenetic characterization.Methods: Breast cancer cell lines MCF7 and SUM159 were cultured in adherent conditions and as mammospheres. Flow cytometry sorting for CD44, CD24 and ALDH was performed. Sorted and unsorted populations, mammospheres and adherent cell cultures were subjected to DNA profiling by array CGH and methylation profiling by Epitect Methyl qPCR array. Methylation status of selected genes was further evaluated by pyrosequencing. Functional impact of methylation was evaluated by mRNA analysis for selected genes.Results: Array CGH did not reveal any genomic differences. In contrast, putative breast cancer stem cells showed altered methylation levels of several genes compared to parental tumor cells.Conclusions: Our results underpin the hypothesis that epigenetic mechanisms seem to play a major role in the regulation of CSCs. However, it is also clear that more efficient methods for CSC enrichment are needed. This work underscores requirement of additional approaches to reveal heterogeneity within breast cancer.

Original languageEnglish
Article number358
JournalBMC Cancer
Volume13
DOIs
StatePublished - Jul 24 2013

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Neoplastic Stem Cells
Epigenomics
Breast Neoplasms
Methylation
Genes
Neoplasms
DNA Fingerprinting
Oligonucleotide Array Sequence Analysis
Flow Cytometry
Cell Culture Techniques
Phenotype
Cell Line
Messenger RNA
Population

ASJC Scopus subject areas

  • Oncology
  • Cancer Research
  • Genetics

Cite this

Balic, M., Schwarzenbacher, D., Stanzer, S., Heitzer, E., Auer, M., Geigl, J. B., ... Dandachi, N. (2013). Genetic and epigenetic analysis of putative breast cancer stem cell models. BMC Cancer, 13, [358]. https://doi.org/10.1186/1471-2407-13-358

Genetic and epigenetic analysis of putative breast cancer stem cell models. / Balic, Marija; Schwarzenbacher, Daniela; Stanzer, Stefanie; Heitzer, Ellen; Auer, Martina; Geigl, Jochen B.; Cote, Richard J; Datar, Ram; Dandachi, Nadia.

In: BMC Cancer, Vol. 13, 358, 24.07.2013.

Research output: Contribution to journalArticle

Balic, M, Schwarzenbacher, D, Stanzer, S, Heitzer, E, Auer, M, Geigl, JB, Cote, RJ, Datar, R & Dandachi, N 2013, 'Genetic and epigenetic analysis of putative breast cancer stem cell models', BMC Cancer, vol. 13, 358. https://doi.org/10.1186/1471-2407-13-358
Balic M, Schwarzenbacher D, Stanzer S, Heitzer E, Auer M, Geigl JB et al. Genetic and epigenetic analysis of putative breast cancer stem cell models. BMC Cancer. 2013 Jul 24;13. 358. https://doi.org/10.1186/1471-2407-13-358
Balic, Marija ; Schwarzenbacher, Daniela ; Stanzer, Stefanie ; Heitzer, Ellen ; Auer, Martina ; Geigl, Jochen B. ; Cote, Richard J ; Datar, Ram ; Dandachi, Nadia. / Genetic and epigenetic analysis of putative breast cancer stem cell models. In: BMC Cancer. 2013 ; Vol. 13.
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