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Gene expression analysis of MCF12A human breast cultures exposed to ethanol or acetaldehyde [miRNA]

ABSTRACT: MCF12A monolayer cultures were exposed to ethanol or acetaldehyde for eiher 1 week or 4 weeks Total RNA samples were assayed for gene expression and for microRNA expression. MicroRNA expression was accomplished using 2 experiments. The first experiment included untreated control, ethanol treated (4 weeks at 2.5 mM) and acetaldehyde treated (4 weeks at 1 mM). The data are presented in order of control1 for the ethanol and acetaldehyde treated samples followed by the ethanol and acetaldehyde treated samples, and then the control2 for the soft agar selected sample followed by the soft agar selected sample. The microRNA names are included in separate columns.

ORGANISM(S): Homo sapiens

PROVIDER: GSE76952 | GEO | 2017/02/10

SECONDARY ACCESSION(S): PRJNA309125

REPOSITORIES: GEO

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Gene expression analysis of MCF12A human breast cultures exposed to ethanol or acetaldehyde [4 weeks]

Project description:Alcoholism is associated with breast cancer incidence and progression, and moderate chronic consumption of ethanol is a risk factor. The mechanisms involved in alcohol's oncogenic effects are unknown, but it has been speculated that they may be mediated by acetaldehyde. Here, we use the immortalized normal human epithelial breast cell line MCF-12A to determine whether short- or long-term exposure to ethanol or to acetaldehyde, using in vivo compatible ethanol concentrations, induces their oncogenic transformation and/or the acquisition of epithelial mesenchymal transition (EMT). Cultures of MCF-12A cells were incubated with 25 mM ethanol or 2.5 mM acetaldehyde for 1 week, or with lower concentrations (1.0-2.5 mM for ethanol, 1.0 mM for acetaldehyde) for 4 weeks. In the 4 wk incubation, cells were also tested for anchorage independence, including isolation of soft agar selected cells (SASC) from the 2.5 mM ethanol incubations. Cells were analyzed by immuno-cytofluorescence, flow cytometry, western blotting, DNA microarrays, RT/PCR, and assays for miRs. We found that short-term exposure to ethanol, but not, in general, to acetaldehyde, was associated with transcriptional upregulation of the metallothionein family genes, alcohol metabolism genes, and genes suggesting the initiation of EMT, but without related phenotypic changes. Long-term exposure to the lower concentrations of ethanol or acetaldehyde induced frank EMT changes in the monolayer cultures and in SASC as demonstrated by changes in cellular phenotype and mRNA expression. This suggests that low concentrations of ethanol, with little or no mediation by acetaldehyde, induce EMT and some traits of oncogenic transformation such as anchorage independence in normal breast epithelial cells. MCF12A monolayer cultures were exposed to ethanol or acetaldehyde for either 1 week or 4 weeks. Total RNA samples were assayed for gene expression and for microRNA expression.

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Project description:RNAseq transcriptomic analysis of MCF10A-CT, MC26 and M1B26 cell lines. MCF10A cells are human not transformed mammary stem-like cells that do not form colonies in soft-agar assay and do not engraft in immunocompromised mice. MC26 are MCF10A derivatives that were submitted to long-term culture in presence of human recombinant BMP2 and IL6 for several weeks until colony formation in soft-agar was detected. M1B26 are MCF10A derivatives that were sorted for high expression of the BMPR1b BMP receptor prior to BMP2 and IL6 treatment and expanded from several soft-agar colonies. M1B26 are more transformed than MC26 based on the frequency of soft-agar colonies forming cells and on the ability to engraft in mice. As a control, MCF10A-CT were kept in culture for similar length of time than MC26 and M1B26 but without BMP2 and IL6 and do not generate soft-agar colonies or engraft in mice. This study aims to document the transcriptomic changes associated with early transformation events mediated by abnormal exposure to BMP2 and inflammatory signal.

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