Genetic and epigenetic analysis of putative breast cancer stem cell models.
Genetic and epigenetic analysis of putative breast cancer stem cell models.
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DOI：
10.1186/1471-2407-13-358
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发表时间：
2013-07-24
期刊：
影响因子：
3.8
通讯作者：
Dandachi N
中科院分区：
文献类型：
--
作者：
Balic M;Schwarzenbacher D;Stanzer S;Heitzer E;Auer M;Geigl JB;Cote RJ;Datar RH;Dandachi N
癌症干细胞模型假设存在一小部分能够维持肿瘤形成、自我更新和分化的肿瘤细胞。在乳腺癌中，发现这些细胞与CD 44 + CD 24-low和ALDH+表型相关。我们的研究是为了评估目前乳腺癌干细胞分析方法的适用性，以通过其广泛的遗传和表观遗传特征来评估乳腺癌细胞的异质性。乳腺癌细胞系MCF 7和SUM 159在贴壁条件下培养并作为乳腺球。对CD 44、CD 24和ALDH进行流式细胞术分选。分选和未分选的群体、乳腺球和贴壁细胞培养物通过阵列CGH进行DNA分析，并通过Epitect Methyl qPCR阵列进行甲基化分析。通过焦磷酸测序进一步评估所选基因的甲基化状态。甲基化的功能影响通过mRNA分析对所选基因进行评价。阵列CGH未显示任何基因组差异。相反，与亲本肿瘤细胞相比，推定的乳腺癌干细胞显示出几个基因的甲基化水平改变。我们的研究结果支持了这样一个假设，即表观遗传机制似乎在CSC的调控中起着重要作用。然而，同样清楚的是，需要更有效的CSC富集方法。这项工作强调了需要额外的方法来揭示乳腺癌的异质性。
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. 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. 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. 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.
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