Unknown,Transcriptomics,Genomics,Proteomics

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MiRNA profiling of MCF-7 and MDA-MB-231 breast cancer cells treated or not with phytoestrogen Emodin

ABSTRACT: Estrogen receptor positive MCF-7 and estrogen receptor negative MDA-MB-231 breast cancer cells treated with 10µg/ml phytoestrogen Emodin (1,3,8-trihydroxy-6-methylanthraquinone) and their control cells (untreated with Emodin) were incubated for 48 hours. All samples were prepared as duplicates to have biological replicates. Isolated RNA samples were used for miRNA microarray analysis. Through this experiment miRNA profiling of breast cancer cells having estrogen receptor and not were revealed upon phytoestrogen Emodin treatment.

ORGANISM(S): Homo sapiens

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PROVIDER: E-MTAB-10170 | biostudies-arrayexpress |

SECONDARY ACCESSION(S): E-MTAB-8310

REPOSITORIES: biostudies-arrayexpress

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Project description:BACKGROUND: The molecular mechanisms resulting in the establishment of regionalized gene expression in the human epididymis involve numerous and complex regulatory elements that have not yet been fully elucidated. Interestingly, more than 200 microRNAs (miRNAs) have been identified in the human epididymis that could be involved in the regulation of mRNA stability and post-transcriptionnal expression in this organ. METHODS: Using a global miRNA microarray approach, we investigated the correlation between the miRNA signature and the gene expression profile found in three distinct regions (caput, corpus and cauda) of human epididymides from 3 donors. In silico prediction of transcript miRNA targets was performed in silico using TargetScan and Miranda software’s. To experimentally verify some of these data, FHCE1 immortalized epididymal cell lines were cotransfected with mimics microRNAs and plasmid constructs containing the 3’UTR of target genes downstream of the luciferase gene. RESULTS: We identified 35 miRNAs differentially expressed between the different segments of the epididymis (fold change > 2, P < 0.01). Among these miRNAs, miR-890, miR-892a, miR-892b, miR-891a, miR-891b belonging to the same cluster located on the X chromosome and specifically expressed in epididymal tissues, are significantly more expressed in the corpus and cauda regions compared to the caput. Interestingly, a strong negative correlation (r= -0,89, P <0.001) was found between the pattern of expression of miR-892b and its potential mRNA target Esrrg (Estrogen Related Receptor Gamma), with miR-145 and Cldn10 mRNA (r= -0,92, P<0.001) and with miR-145 and Cftr mRNA (r= -0,69, P<0.05). We confirmed that miR-145 and miR-892b inhibit the expression of the luciferase reporter via Cldn10 and Esrrg 3’ UTRs respectively, whereas miR-145 does not affect the expression of luciferase after its co-transfection with the plasmid containing the Cftr 3’UTR. CONCLUSION: Overall, our study shows that the expression of miRNAs is segmented along the different regions of the human epididymal duct and correlates with the pattern of target gene expression in these regions. Therefore, epididymal miRNAs may be in charge of the maintenance of gene expression profile in the epididymis that dictate segment-specific secretion of proteins and establish physiological compartments that directly or indirectly affect sperm maturation and fertility. Normal human epididymides from three donors of 26–50 years of age, were collected and dissected into 3 anatomical regions (caput, corpus and cauda).A cross-comparison between the miRNA signature found in different segments was performed using GeneChip miRNA Array 1.0 (Affimetrix, Santa Clara, CA).

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MiRNA profiling of MCF-7 and MDA-MB-231 breast cancer cells treated or not with phytoestrogen Emodin

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