Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells.
Adult
Aromatase
/ genetics
Cell Line, Tumor
Cumulus Cells
/ physiology
Female
Follicle Stimulating Hormone
/ metabolism
Gene Expression Profiling
Gene Expression Regulation, Developmental
Humans
Leukemia Inhibitory Factor Receptor alpha Subunit
/ genetics
MicroRNAs
/ genetics
Oligonucleotide Array Sequence Analysis
Ovarian Follicle
/ cytology
PTEN Phosphohydrolase
/ genetics
Primary Cell Culture
Receptors, FSH
/ genetics
Young Adult
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
10 02 2020
10 02 2020
Historique:
received:
16
09
2019
accepted:
08
01
2020
entrez:
12
2
2020
pubmed:
12
2
2020
medline:
21
11
2020
Statut:
epublish
Résumé
MicroRNAs (miRNAs) are known post-transcriptional regulators of various biological processes including ovarian follicle development. We have previously identified miRNAs from human pre-ovulatory ovarian granulosa cells that are expressed from the intronic regions of two key genes in normal follicular development: FSH receptor (FSHR) and CYP19A1, the latter encoding the aromatase enzyme. The present study aims to identify the target genes regulated by these miRNAs: hsa-miR-548ba and hsa-miR-7973, respectively. The miRNAs of interest were transfected into KGN cell line and the gene expression changes were analyzed by Affymetrix microarray. Potential miRNA-regulated genes were further filtered by bioinformatic target prediction algorithms and validated for direct miRNA:mRNA binding by luciferase reporter assay. LIFR, PTEN, NEO1 and SP110 were confirmed as targets for hsa-miR-548ba. Hsa-miR-7973 target genes ADAM19, PXDN and FMNL3 also passed all verification steps. Additionally, the expression pattern of the miRNAs was studied in human primary cumulus granulosa cell culture in relation to the expression of their host genes and FSH stimulation. Based on our findings we propose the involvement of hsa-miR-548ba in the regulation of follicle growth and activation via LIFR and PTEN. Hsa-miR-7973 may be implicated in the modulation of extracellular matrix and cell-cell interactions by regulating the expression of its identified targets.
Identifiants
pubmed: 32042028
doi: 10.1038/s41598-020-59186-x
pii: 10.1038/s41598-020-59186-x
pmc: PMC7010774
doi:
Substances chimiques
FSHR protein, human
0
LIFR protein, human
0
Leukemia Inhibitory Factor Receptor alpha Subunit
0
MIRN548 microRNA, human
0
MIRN548BA microRNA, human
0
MIRN7973-1 microRNA, human
0
MicroRNAs
0
Receptors, FSH
0
Follicle Stimulating Hormone
9002-68-0
Aromatase
EC 1.14.14.1
CYP19A1 protein, human
EC 1.14.14.1
PTEN Phosphohydrolase
EC 3.1.3.67
PTEN protein, human
EC 3.1.3.67
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2300Références
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