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Title: Essential role of miR-200c in regulating self-renewal of breast cancer stem cells initiating from the counter parts of mammary epithelium      
dateReleased:
04-26-2015
description:
To explore the roles of essential miRNAs in regulating self-renewal of breast cancer stem cells (BCSCs), which initiate from mammary epithelial stem cells (MaSCs). CD44+CD24-/low cells and MUC1-ESA+ cells were isolated by fluorescence-activated cell sorting (FACS) from breast cancer cell line MCF-7 and normal mammary epithelial cell line MCF-10A, and were verified as BCSCs and MaSCs by clonogenic assay and multipotential differentiation experiment in 2-dimensional (2-D) and 3-D cultures, respectively. Using microarray containing oligonucleotides corresponding to 509 miRNAs from human, mouse, and rat genomes. We obtained candidate miRNAs in regulating breast tumorigenesis. One representative miRNA (miR-200c) was proved to regulate stemness of BCSCs and MaSCs in vitro and in vivo by miR-200c agomir transfection. We validated that miR-200c negatively regulated PDCD10, an apoptosis regulator, in BCSCs and MaSCs. Our miRNA microarray includes 509 mature miRNA sequences were downloaded from the miRNA Registry. We designed eight short oligos possessing no homology with all existed RNA sequence and produced their corresponding synthetic miRNAs by in vitro transcription using Ambion’s miRNA Probe Construction kit. All miRNA probe sequences were designed to be complementary to the full-length mature miRNA. The probe sequence were concatenated up to 40 nt and modified with 5 0 -amino-modifier C6. Oligonucleotide probes were synthesized and dissolved in EasyArray TM spotting solution at 40 mM concentration. Each probe was printed in triplicate using a SmartArray TM microarrayer. Arrays were scanned with a confocal LuxScan TM scanner (CapitalBio Corp.)Significance Analysis of Microarrays(SAM, version 2.1) was performed using two class-unpaired comparison in the SAM procedure.
privacy:
not applicable
aggregation:
instance of dataset
ID:
E-GEOD-68271
refinement:
raw
alternateIdentifiers:
68271
keywords:
functional genomics
dateModified:
05-02-2015
availability:
available
types:
gene expression
name:
Homo sapiens
ID:
A-GEOD-4508
name:
Tsinghua University mammalian 2K microRNA microarray
accessURL: https://www.ebi.ac.uk/arrayexpress/files/E-GEOD-68271/E-GEOD-68271.raw.1.zip
storedIn:
ArrayExpress
qualifier:
gzip compressed
format:
TXT
accessType:
download
authentication:
none
authorization:
none
accessURL: https://www.ebi.ac.uk/arrayexpress/files/E-GEOD-68271/E-GEOD-68271.processed.1.zip
storedIn:
ArrayExpress
qualifier:
gzip compressed
format:
TXT
accessType:
download
authentication:
none
authorization:
none
accessURL: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE68271
storedIn:
Gene Expression Omnibus
qualifier:
not compressed
format:
HTML
accessType:
landing page
primary:
true
authentication:
none
authorization:
none
abbreviation:
EBI
homePage: http://www.ebi.ac.uk/
ID:
SCR:004727
name:
European Bioinformatics Institute
homePage: https://www.ebi.ac.uk/arrayexpress/
ID:
SCR:002964
name:
ArrayExpress
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