MicroRNA-183-96-182 Cluster Regulates Bovine Granulosa Cell Proliferation and Cell Cycle Transition by Coordinately Targeting FOXO1.
Large-scale expression profiling of micro-RNAs (miRNAs) in bovine granulosa cells from dominant and subordinate follicles on Day 19 of the estrous cycle revealed enriched micro-RNA-183-96-182 cluster miRNAs in preovulatory dominant follicles that coordinately regulate the forkhead box protein O1 (FOXO1) gene. However, little is known about the role of this cluster in bovine granulosa cell function. We used an in vitro granulosa cell culture model to investigate this role. Granulosa cells aspirated from small growing follicles (3-5 mm in diameter) were cultured in Dulbecco modified Eagle medium/F-12 medium supplemented with fetal bovine serum and transfected with locked nucleic acid-based miRNA mimics, inhibitors, and corresponding negative controls. Overexpression of the miRNA cluster resulted in suppression of FOXO1 mRNA and protein, whereas inhibition of the cluster increased expression of FOXO1 mRNA. Overexpression also increased the relative rate of cell proliferation, whereas inhibition slowed it down. Similarly, the proportion of cells under G0/G1 arrest declined, whereas the ratio of cells in S phase increased in response to miR-183-96-182 overexpression. Selective knockdown of FOXO1 mRNA using anti-FOXO1 small interfering RNA increased the rate of granulosa cell proliferation, decreased the proportion of cells under G0/G1 arrest, and increased the proportion of cells in the S phase of cell cycle. Our data suggest that miR-183-96-182 cluster miRNAs promote proliferation and G1/S transition of bovine granulosa cells by coordinately targeting FOXO1, suggesting a critical role in granulosa cell function. MicroRNA-183-96-182 cluster regulates bovine granulosa cell function by targeting FOXO1 gene.
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany.
Samuel
S
Gebremedhn
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany.
Dessie
D
Salilew-Wondim
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany Teaching and Research Station Frankenforst, Faculty of Agriculture, University of Bonn, Königswinter, Germany Center of Integrated Dairy Research, University of Bonn, Bonn, Germany.
Michael
M
Hoelker
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany Teaching and Research Station Frankenforst, Faculty of Agriculture, University of Bonn, Königswinter, Germany.
Franca
F
Rings
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany.
Christiane
C
Neuhoff
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany.
Ernst
E
Tholen
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany Center of Integrated Dairy Research, University of Bonn, Bonn, Germany.
Karl
K
Schellander
Institute of Animal Science, Department of Animal Breeding and Husbandry, University of Bonn, Bonn, Germany Center of Integrated Dairy Research, University of Bonn, Bonn, Germany tesfaye@itw.uni-bonn.de.
Dawit
D
Tesfaye
D000818:Animals; D002417:Cattle; D002453:Cell Cycle; D049109:Cell Proliferation; D005260:Female; D000071161:Forkhead Box Protein O1 / Q000235:genetics / Q000378:metabolism*; D006107:Granulosa Cells / Q000502:physiology*; D035683:MicroRNAs / Q000378:metabolism*
D000071161:Forkhead Box Protein O1; D035683:MicroRNAs
FOXO1; cell cycle; cell proliferation; granulosa cell; miR-183-96-182 cluster