title: AMFR gene silencing inhibits the differentiation of porcine preadipocytes.
abstract: Our study clarifies the role of the autocrine motility factor receptor (AMFR) gene in porcine preadipocyte differentiation. AMFR-siRNA was transfected into porcine preadipocytes and the preadipocytes were induced to differentiation. Subsequently, qRT-PCR was conducted to examine changes in mRNA expression of a series of genes in porcine preadipocytes, including AMFR, sterol-regulatory element-binding protein-1a (SREBP1a), SREBP2, insulin-induced gene 1 (Insig1), and Insig2. Expression changes in the mRNA of genes regulating adipocyte differentiation were also analyzed using qRT-PCR, including peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBPα), and Kruppel-like factor 2 (KLF2). Western blot analysis was conducted to examine the changes in AMFR protein expression in porcine preadipocytes. Additionally, morphological changes in differentiated porcine preadipocytes were examined by oil red O staining, and changes in optical density (OD) values were measured using an ultraviolet spectrophotometer. At 24 h after transfection with AMFR-siRNA, AMFR mRNA expression significantly reduced (P < 0.01), and AMFR protein expression markedly decreased (P < 0.05). The mRNA expression of SREBP1a, SREBP2, Insig1, and C/EBPα was significantly reduced (P < 0.01), whereas the expression of KLF2 mRNA was significantly elevated (P < 0.01). After induction of preadipocyte differentiation, the number of lipid droplets decreased in the AMFR-silenced group, and the OD value markedly reduced (P < 0.05). In addition, the expression of C/EBPα mRNA significantly decreased (P < 0.05), whereas the expression of KLF2 mRNA considerably increased (P < 0.05). Taken together, silencing of the AMFR gene inhibits the differentiation of porcine preadipocytes.
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: C Z
authors.initials: CZ
authors.lastname: Chen
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Y N
authors.initials: YN
authors.lastname: Zhu
authors.affiliation: College of Animal Science and Technology, Jilin Agriculture University, Changchun, China.
authors.forename: M L
authors.initials: ML
authors.lastname: Chai
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: L S
authors.initials: LS
authors.lastname: Dai
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: H
authors.initials: H
authors.lastname: Jiang
authors.affiliation: College of Animal Science and Technology, Jilin Agricultural Science and Technology College, Jilin, China.
authors.forename: L J
authors.initials: LJ
authors.lastname: Zhang
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: S Y
authors.initials: SY
authors.lastname: Liu
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Q Y
authors.initials: QY
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Jilin Agriculture University, Changchun, China.
authors.forename: W F
authors.initials: WF
authors.lastname: Lu
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.|State & Local Joint Engineering Laboratory for Animal Models of Human Diseases, Jilin University, Changchun, China.
authors.forename: J B
authors.initials: JB
authors.lastname: Zhang
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D020868:Gene Silencing; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D060285:Receptors, Autocrine Motility Factor / Q000235:genetics / Q000378:metabolism*; D051780:Sterol Regulatory Element Binding Protein 1 / Q000235:genetics / Q000378:metabolism; D051782:Sterol Regulatory Element Binding Protein 2 / Q000235:genetics / Q000378:metabolism; D013552:Swine
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; D051741:Kruppel-Like Transcription Factors; D047495:PPAR gamma; D051780:Sterol Regulatory Element Binding Protein 1; D051782:Sterol Regulatory Element Binding Protein 2; D060285:Receptors, Autocrine Motility Factor