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Let-7g induces granulosa cell apoptosis by targeting MAP3K1 in the porcine ovary |
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| Institution: | 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 |
| 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. |
| 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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