Let-7g induces granulosa cell apoptosis by targeting MAP3K1 in the porcine ovary |
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| Affiliation: | 1. Department of Biomaterials, Radboud University Medical Center, Nijmegen, The Netherlands;2. Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands;3. Department of Biophysics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Centre, Nijmegen, The Netherlands;4. Department of Otorhinolaryngology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, Nijmegen, The Netherlands;1. Animal Science and Technology College, Beijing University of Agriculture, Beijing, China;2. College of Animal Science and Technology, China Agricultural University, Beijing, China;1. Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium;2. National Institute of Livestock and Grassland Science, Tsukuba, Ibaraki, Japan;3. Transgenic Pig Research Unit, National Institute of Agrobiological Sciences, Ibaraki, Japan;4. NARIC-Agricultural Biotechnology Institute, Szent István University, Godollo, Hungary;5. Genetic Diversity Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan;1. Laboratorio de Biología y Quimioterapia de Parasitosis Tropicales, Área de Salud, Instituto de Estudios Avanzados (IDEA), Caracas, Bolivarian Republic of Venezuela;2. Laboratorio de Cultivo de Tejidos y Biología de Tumores, Instituto de Biología Experimental (IBE), Universidad Central de Venezuela, Caracas, Bolivarian Republic of Venezuela;3. Laboratorio de Ingeniería de Tejidos Humanos, Área de Salud, Instituto de Estudios Avanzados (IDEA), Caracas, Bolivarian Republic of Venezuela;4. Laboratorio de Pesquisa Selectiva, Área de Salud, Instituto de Estudios Avanzados (IDEA), Caracas, Bolivarian Republic of Venezuela;5. Laboratorio de Fitoquímica, Área de Agricultura y Soberanía Alimentaria, Instituto de Estudios Avanzados (IDEA), Caracas, Bolivarian Republic of Venezuela;6. Laboratorio de Síntesis Orgánica y Compuestos Naturales, Centro de Química, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Bolivarian Republic of Venezuela;7. Laboratorio de Química Orgánica y Biomolecular, CMN, Universidad Industrial de Santander, Parque Tecnológico Guatiguará, Km 2 Vía Refugio, Piedecuesta, 681011, Colombia;1. Department of Environmental Health and University of Cincinnati, College of Medicine, United States;2. Department of Ophthalmology, University of Cincinnati, College of Medicine, United States;3. Division of Developmental Biology, Cincinnati Children׳s Hospital Medical Center, United States |
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| Abstract: | Follicular atresia mainly results from apoptosis of granulosa cells (GCs). Our previous microRNA array data indicated that the miRNA let-7g level increases significantly during porcine ovary follicular atresia. It is uncertain if GCs apoptosis is mediated by microRNA let-7g. In this study, the expression levels of the apoptosis-associated genes CASP3, BAX and BIM were significantly upregulated when let-7g mimic was transfected into porcine GCs, and the anti-apoptotic genes BCL-2 and MCL-1 were significantly downregulated. The apoptosis rate was measured by flow cytometry, and our results indicated that let-7g significantly enhanced GCs apoptosis. In further studies, we found that overexpression of let-7g induced the expression of FoxO1 in GCs and led to nuclear accumulation of dephosphorylated FoxO1. In addition, the effect of let-7g on FoxO1 expression and dephosphorylation resulted from repression of the expression of the MAP3K1 gene in porcine GCs. The site on MAP3K1 mRNA targeted by let-7g was confirmed by luciferase reporter assay. The anti-apoptotic effect of MAP3K1 was validated by silencing MAP3K1 using small interfering RNA technology. In conclusion, our data indicate that let-7g induces porcine GCs apoptosis by inhibiting the MAP3K1 gene, which promotes FoxO1 expression and dephosphorylation with nuclear accumulation. |
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| Keywords: | Let-7g MAP3K1 FoxO1 Granulosa cell Apoptosis miRNAs" },{" #name" :" keyword" ," $" :{" id" :" kw0035" }," $$" :[{" #name" :" text" ," _" :" mircoRNAs 3′ UTR" },{" #name" :" keyword" ," $" :{" id" :" kw0045" }," $$" :[{" #name" :" text" ," _" :" 3′-untranslated regions GCs" },{" #name" :" keyword" ," $" :{" id" :" kw0055" }," $$" :[{" #name" :" text" ," _" :" granulosa cells PTEN" },{" #name" :" keyword" ," $" :{" id" :" kw0065" }," $$" :[{" #name" :" text" ," _" :" phosphatase and tensin homolog PBS" },{" #name" :" keyword" ," $" :{" id" :" kw0075" }," $$" :[{" #name" :" text" ," _" :" phosphate buffered saline PI" },{" #name" :" keyword" ," $" :{" id" :" kw0085" }," $$" :[{" #name" :" text" ," _" :" propidium iodide BSA" },{" #name" :" keyword" ," $" :{" id" :" kw0095" }," $$" :[{" #name" :" text" ," _" :" bovine serum albumin DAPI" },{" #name" :" keyword" ," $" :{" id" :" kw0105" }," $$" :[{" #name" :" text" ," _" :" AT specific fluorescent dye 4′-6-diamidino-2-phenylindole RIPA" },{" #name" :" keyword" ," $" :{" id" :" kw0115" }," $$" :[{" #name" :" text" ," _" :" radio-immunoprecipitation assay TBST" },{" #name" :" keyword" ," $" :{" id" :" kw0125" }," $$" :[{" #name" :" text" ," _" :" tris buffered saline and tween 20 GO" },{" #name" :" keyword" ," $" :{" id" :" kw0135" }," $$" :[{" #name" :" text" ," _" :" gene ontology KEGG" },{" #name" :" keyword" ," $" :{" id" :" kw0145" }," $$" :[{" #name" :" text" ," _" :" Kyoto encyclopedia of genes and genomes FASL" },{" #name" :" keyword" ," $" :{" id" :" kw0155" }," $$" :[{" #name" :" text" ," _" :" fatty acid synthase ligand BAX" },{" #name" :" keyword" ," $" :{" id" :" kw0165" }," $$" :[{" #name" :" text" ," _" :" Bcl2-associated X protein BCL2" },{" #name" :" keyword" ," $" :{" id" :" kw0175" }," $$" :[{" #name" :" text" ," _" :" B-cell CLL/lymphoma 2 MCL1" },{" #name" :" keyword" ," $" :{" id" :" kw0185" }," $$" :[{" #name" :" text" ," _" :" myeloid cell leukemia sequence 1 FoxO1" },{" #name" :" keyword" ," $" :{" id" :" kw0195" }," $$" :[{" #name" :" text" ," _" :" Forkhead box O 1 BIM" },{" #name" :" keyword" ," $" :{" id" :" kw0205" }," $$" :[{" #name" :" text" ," _" :" Bcl2-Interacting Mediator of Cell Death TRAIL" },{" #name" :" keyword" ," $" :{" id" :" kw0215" }," $$" :[{" #name" :" text" ," _" :" tumor necrosis factor-related apoptosis-inducing ligand WT" },{" #name" :" keyword" ," $" :{" id" :" kw0225" }," $$" :[{" #name" :" text" ," _" :" wild-type pmirGLO-MAP3K1-3′UTR vector Mutant" },{" #name" :" keyword" ," $" :{" id" :" kw0235" }," $$" :[{" #name" :" text" ," _" :" mutant pmirGLO-MAP3K1-3′UTR vector MAP3K1" },{" #name" :" keyword" ," $" :{" id" :" kw0245" }," $$" :[{" #name" :" text" ," _" :" Mitogen-Activated Protein Kinase Kinase Kinase 1,E3 Ubiquitin Protein JNK" },{" #name" :" keyword" ," $" :{" id" :" kw0255" }," $$" :[{" #name" :" text" ," _" :" c-Jun NH2-terminal kinase p38-MAPK" },{" #name" :" keyword" ," $" :{" id" :" kw0265" }," $$" :[{" #name" :" text" ," _" :" p38 mitogen-activated protein kinases ERK" },{" #name" :" keyword" ," $" :{" id" :" kw0275" }," $$" :[{" #name" :" text" ," _" :" extracellular signal-regulated protein kinase NF-kappa-B" },{" #name" :" keyword" ," $" :{" id" :" kw0285" }," $$" :[{" #name" :" text" ," _" :" nuclear factor-kappa B |
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