MicroRNA as Crucial Regulators of Gene Expression in Estradiol-Treated Human Endothelial Cells.
BACKGROUND/AIMS
Estrogen signalling plays an important role in vascular biology as it modulates vasoactive and metabolic pathways in endothelial cells. Growing evidence has also established microRNA (miRNA) as key regulators of endothelial function. Nonetheless, the role of estrogen regulation on miRNA profile in endothelial cells is poorly understood. In this study, we aimed to determine how estrogen modulates miRNA profile in human endothelial cells and to explore the role of the different estrogen receptors (ERα, ERβ and GPER) in the regulation of miRNA expression by estrogen.


METHODS
We used miRNA microarrays to determine global miRNA expression in human umbilical vein endothelial cells (HUVEC) exposed to a physiological concentration of estradiol (E2; 1 nmol/L) for 24 hours. miRNA-gene interactions were computationally predicted using Ingenuity Pathway Analysis and changes in miRNA levels were validated by qRT-PCR. Role of ER in the E2-induced miRNA was additionally confirmed by using specific ER agonists and antagonists.


RESULTS
miRNA array revealed that expression of 114 miRNA were significantly modified after E2 exposition. Further biological pathway analysis revealed cell death and survival, lipid metabolism, reproductive system function, as the top functions regulated by E2. We validated changes in the most significantly increased (miR-30b-5p, miR-487a-5p, miR-4710, miR-501-3p) and decreased (miR-378h and miR-1244) miRNA and the role of ER in these E2-induced miRNA was determined. Results showed that both classical, ERα and ERβ, and membrane-bound ER, GPER, differentially regulated specific miRNA. In silico analysis of validated miRNA promoters identified specific ER binding sites.


CONCLUSION
Our findings identify differentially expressed miRNA pathways linked to E2 in human endothelial cells through ER, and provide new insights by which estrogen can modulate endothelial function.
['Vidal-Gómez|Xavier|X|', 'Pérez-Cremades|Daniel|D|', 'Mompeón|Ana|A|', 'Dantas|Ana Paula|AP|', 'Novella|Susana|S|', 'Hermenegildo|Carlos|C|']
D002478:Cells, Cultured D016000:Cluster Analysis D015536:Down-Regulation D004958:Estradiol D020869:Gene Expression Profiling D061307:Human Umbilical Vein Endothelial Cells D006801:Humans D035683:MicroRNAs D020411:Oligonucleotide Array Sequence Analysis D025341:Principal Component Analysis D011960:Receptors, Estrogen D043562:Receptors, G-Protein-Coupled D015854:Up-Regulation
Endothelial cells MiRNA Estrogen receptors Estradiol Epigenetic regulation