Unknown,Transcriptomics,Genomics,Proteomics

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The effect of miRNA overexpression on gene expression profile of MCF-7 cells


ABSTRACT: To identify microRNAs impacting estrogen receptor ERα expression in breast cancer, we have screened ER-positive breast cancer cells with a library of pre-miRs, and systematically monitored the ERα expression by protein lysate microarrays. There was a significant enrichment of the in silico predicted ERα targeting microRNAs among the hits. The most potent pre-miRs miR-18a/b, miR-193b, miR-206, and miR-302c, were confirmed to directly target ERα and to repress estrogen-responsive genes. The effect of miRNA overexpression on gene expression profile of MCF-7 cells was studied. Furthermore, miR-18a and miR-18b showed increased expression in ERα-negative as compared to ERα-positive clinical tumors. In summary, we present systematic and direct functional and correlative clinical evidence on microRNAs inhibiting ERα signaling in breast cancer. MCF-7 cells (300 000 per well on 6-well plates) were transfected with an siRNA for ERα or with Ambion pre-miR� constructs for miR-18a, miR-193b, miR-206, miR-302c, or pre-miR negative control #1 (scrambled pre-miR) at 20 nM, and incubated for 24h.

ORGANISM(S): Homo sapiens

SUBMITTER: Suvi-Katri Leivonen 

PROVIDER: E-GEOD-14847 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Protein lysate microarray analysis to identify microRNAs regulating estrogen receptor signaling in breast cancer cell lines.

Leivonen S-K SK   Mäkelä R R   Ostling P P   Kohonen P P   Haapa-Paananen S S   Kleivi K K   Enerly E E   Aakula A A   Hellström K K   Sahlberg N N   Kristensen V N VN   Børresen-Dale A-L AL   Saviranta P P   Perälä M M   Kallioniemi O O  

Oncogene 20090817 44


Predicting the impact of microRNAs (miRNAs) on target proteins is challenging because of their different regulatory effects at the transcriptional and translational levels. In this study, we applied a novel protein lysate microarray (LMA) technology to systematically monitor for target protein levels after high-throughput transfections of 319 pre-miRs into breast cancer cells. We identified 21 miRNAs that downregulated the estrogen receptor-alpha (ERalpha), as validated by western blotting and q  ...[more]

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