Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells

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http://hdl.handle.net/10138/319157

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Rooda , I , Hensen , K , Kaselt , B , Kasvandik , S , Pook , M , Kurg , A , Salumets , A & Velthut-Meikas , A 2020 , ' Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells ' , Scientific Reports , vol. 10 , no. 1 , 2300 . https://doi.org/10.1038/s41598-020-59186-x

Title: Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells
Author: Rooda, Ilmatar; Hensen, Kati; Kaselt, Birgitta; Kasvandik, Sergo; Pook, Martin; Kurg, Ants; Salumets, Andres; Velthut-Meikas, Agne
Contributor: University of Helsinki, HUS Gynecology and Obstetrics
Date: 2020-02-10
Language: eng
Number of pages: 13
Belongs to series: Scientific Reports
ISSN: 2045-2322
URI: http://hdl.handle.net/10138/319157
Abstract: MicroRNAs (miRNAs) are known post-transcriptional regulators of various biological processes including ovarian follicle development. We have previously identified miRNAs from human preovulatory 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.
Subject: LEUKEMIA INHIBITORY FACTOR
UP-REGULATION
RECEPTOR
PROLIFERATION
ACTIVATION
PROMOTES
CUMULUS
MARKERS
PATHWAY
OOCYTE
318 Medical biotechnology
3111 Biomedicine
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