miR-106b-5p靶向KLF4调控山羊肌内前体脂肪细胞分化

doi:10.11843/j.issn.0366-6964.2020.006

Abstract

Abstract: This study aimed to explore the effect of miR-106b-5p on the differentiation of goat intramuscular preadipocytes, and to confirm whether miR-106b-5p played its roles via targeting KLF4. The quantitative real-time PCR (qRT-PCR) was used to detect the expression pattern of miR-106b-5p during the differentiation of goat intramuscular preadipocytes. The miR-106b-5p mimic and miR-106b-5p inhibitor were transfected into goat intramuscular preadipocytes cultured in vitro by liposome transfection. The effect of miR-106b-5p on lipid droplet accumulation was verified using the Oil Red O Staining. The expression levels of predicted target gene KLF4 targeting miR-106b-5p and adipogenic marker genes were detected using qRT-PCR. The target relationship between miR-106b-5p and KLF4 was detected by dual luciferase reporter system. The result of qRT-PCR showed that the highest expression level of miR-106b-5p was at day 3 of induced differentiation of goat intramuscular preadipocytes. The Oil Red O staining result showed that lipid droplet accumulation was decreased after treated by miR-106b-5p inhibitor, however, it had an opposite result after treated by miR-106b-5p mimic in goat intramuscular adipocytes. The expression level of PPARγ was significantly decreased (P<0.05) and the expression level of KLF4 was extremely significantly increased (P<0.01) in the miR-106b-5p inhibitor treatment. The expression levels of LPL and PPARγ were extremely significantly increased (P<0.01) in the miR-106b-5p mimic treatment. The luciferase activity assay result showed that luciferase activity of KLF4 was significantly inhibited in the miR-106b-5p mimic treatment. miR-106b-5p promotes the differentiation of goat intramuscular preadipocytes via targeting and negatively regulating the expression of KLF4.

Key words: goat, miR-106b-5p, KLF4, adipocyte, luciferase activity assay

CLC Number:

S827.2

DU Yu, ZHAO Yue, LIN Yaqiu, ZHU Jiangjiang, WANG Yong, MA Jieqiong, XIE Guangjie. miR-106b-5p Regulates the Differentiation of Goat Intramuscular Preadipocytes by Targeting KLF4[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1219-1228.

References 36

[1]	ZHOU P,XU W Y,PENG X L,et al.Large-scale screens of miRNA-mRNA interactions unveiled that the 3'UTR of a gene is targeted by multiple miRNAs[J].PLoS One,2013,8(7):e68204.
[2]	CHI S W,ZANG J B,MELE A,et al.Argonaute HITS-CLIP decodes microRNA-mRNA interaction maps[J].Nature, 2009, 460(7254):479-486.
[3]	HAFNER M,LANDTHALER M,BURGER L,et al.Transcriptome-wide identification of RNA-binding protein and microRNA target sites by PAR-CLIP[J].Cell,2010,141(1):129-141.
[4]	张莹姣,乔利英,景炅婕,等.miR-194-5p靶向调控BCKDHA基因的表达[J].畜牧兽医学报,2017,48(8):1416-1423.ZHANG Y J,QIAO L Y,JING J J,et al.miR-194-5p targets BCKDHA gene to regulate its expression[J].Acta Veterinaria et Zootechnica Sinica,2017,48(8):1416-1423.(in Chinese)
[5]	SHI C M,ZHANG M,TONG M L,et al.miR-148a is associated with obesity and modulates adipocyte differentiation of mesenchymal stem cells through Wnt signaling[J].Sci Rep,2015,5:9930.
[6]	CHEN L,DAI Y M,JI C B,et al.MiR-146b is a regulator of human visceral preadipocyte proliferation and differentiation and its expression is altered in human obesity[J].Mol Cell Endocrinol,2014,393(1-2):65-74.
[7]	MA L,QIU H L,CHEN Z,et al.miR-25 modulates triacylglycerol and lipid accumulation in goat mammary epithelial cells by repressing PGC-1beta[J].J Anim Sci Biotechnol,2018,9:48.
[8]	ZHANG Y Y,WANG Y N,WANG H B,et al.MicroRNA-224 impairs adipogenic differentiation of bovine preadipocytes by targeting LPL[J].Mol Cell Probes,2019,44:29-36.
[9]	NI S J,WENG W W,XU M D,et al.miR-106b-5p inhibits the invasion and metastasis of colorectal cancer by targeting CTSA[J].Onco Targets Ther,2018,11:3835-3845.
[10]	ZHUANG M,ZHAO S L,JIANG Z,et al.MALAT1 sponges miR-106b-5p to promote the invasion and metastasis of colorectal cancer via SLAIN2 enhanced microtubules mobility[J].EBioMedicine,2019,41:286-298.
[11]	REN J Y,ZHANG J,XU N,et al.Signature of circulating microRNAs as potential biomarkers in vulnerable coronary artery disease[J].PLoS One,2013,8(12):e80738.
[12]	张静,李素芳,陈红,等.miR-106b-5p在调节内皮细胞基因表达谱中的作用[J].北京大学学报:医学版, 2019, 51(2):221-227.ZHANG J,LI S F,CHEN H,et al.Role of miR-106b-5p in the regulation of gene profiles in endothelial cells[J].Journal of Peking University:Health Sciences,2019,51(2):221-227.(in Chinese)
[13]	TSUKITA S,YAMADA T,TAKAHASHI K,et al.MicroRNAs 106b and 222 improve hyperglycemia in a mouse model of insulin-deficient diabetes via pancreatic β-Cell Proliferation[J].EBioMedicine,2017,15:163-172.
[14]	WANG Q,LI Y C,WANG J H,et al.miR-17-92 cluster accelerates adipocyte differentiation by negatively regulating tumor-suppressor Rb2/p130[J].Proc Natl Acad Sci U S A,2008,105(8):2889-2894.
[15]	SHI C M,HUANG F Y,GU X H,et al.Adipogenic miRNA and meta-signature miRNAs involved in human adipocyte differentiation and obesity[J].Oncotarget,2016,7(26):40830-40845.
[16]	YE D,LOU G H,ZHANG T B,et al.MiR-17 family-mediated regulation of Pknox1 influences hepatic steatosis and insulin signaling[J].J Cell Mol Med,2018,22(12):6167-6175.
[17]	KROL J,LOEDIGE I,FILIPOWICZ W.The widespread regulation of microRNA biogenesis,function and decay[J].Nat Rev Genet,2010,11(9):597-610.
[18]	SON Y H,KA S,KIM A Y,et al.Regulation of adipocyte differentiation via MicroRNAs[J].Endocrinol Metab (Seoul),2014, 29(2):122-135.
[19]	HA M,KIM V N.Regulation of microRNA biogenesis[J].Nat Rev Mol Cell Biol,2014,15(8):509-524.
[20]	LEUNG A K L,SHARP P A.Function and localization of microRNAs in mammalian cells[J].Cold Spring Harb Symp Quant Biol,2006,71:29-38.
[21]	赵艳艳.miR-142和miR-144靶向FoxO1基因调节绵羊前体脂肪细胞分化的研究[D].晋中:山西农业大学,2018.ZHAO Y Y.Study on miR-142 and miR-144 down-regulation contributions to differentiation of ovine preadipocytes by targeting FoxO1 gene[D].Jinzhong:Shanxi Agricultural University,2018.(in Chinese)
[22]	张莹姣.MiR-194-5p靶向BCKDHA基因调控绵羊脂肪生成机制的研究[D].晋中:山西农业大学,2017.ZHANG Y J.Studies on the regulation mechanism of MiR-194-5p by targeting BCKDHA gene in ovine adipogenesis[D]. Jinzhong:Shanxi Agricultural University,2017.(in Chinese)
[23]	LING H Y,WEN G B,FENG S D,et al.MicroRNA-375 promotes 3T3-L1 adipocyte differentiation through modulation of extracellular signal-regulated kinase signalling[J].Clin Exp Pharmacol Physiol,2011,38(4):239-246.
[24]	QI R L,WANG J,WANG Q,et al.MicroRNA-425 controls lipogenesis and lipolysis in adipocytes[J].Biochim Biophys Acta Mol Cell Biol Lipids,2019,1864(5):744-755.
[25]	LI M H,LIU Z J,ZHANG Z Z,et al.miR-103 promotes 3T3-L1 cell adipogenesis through AKT/mTOR signal pathway with its target being MEF2D[J].Biol Chem,2015,396(3):235-244.
[26]	SHI X E,LI Y F,JIA L,et al.MicroRNA-199a-5p affects porcine preadipocyte proliferation and differentiation[J].Int J Mol Sci, 2014, 15(5):8526-8538.
[27]	CHEN C,DENG Y,HU X G,et al.miR-128-3p regulates 3T3-L1 adipogenesis and lipolysis by targeting Pparg and Sertad2[J].J Physiol Biochem,2018,74(3):381-393.
[28]	YU L X,ZHANG B L,YANG M Y,et al.MicroRNA-106b-5p promotes hepatocellular carcinoma development via modulating FOG2[J].Onco Targets Ther,2019,12:5639-5647.
[29]	林森.KLF4对山羊肌内前体脂肪细胞分化的调控作用[D].成都:西南民族大学,2018.LIN S.Regulation of KLF4 on intramuscular preadipocyte differentiation in goat[D].Chengdu:Southwest Minzu University, 2018.(in Chinese)
[30]	JENO Y G,LEE J H,JI Y,et al.RNF20 functions as a transcriptional coactivator for PPARγ by promoting NCoR1 degradation in adipocytes[J].Diabetes,2020,69(1):20-34.
[31]	马雪瑶.Bta-miR-130调控秦川牛前体脂肪细胞分化的作用机制研究[D].杨凌:西北农林科技大学,2019.MA X Y.The molecular mechanism of Bta-MiR-130 regulates preadipocyte differentiation in Qinchuan beef cattle[D]. Yangling:Northwest A&F University,2019.(in Chinese)
[32]	SHI H B,LUO J,ZHU J J,et al.PPARγ regulates genes involved in triacylglycerol synthesis and secretion in mammary gland epithelial cells of dairy goats[J].PPAR Res,2013,2013:310948.
[33]	TONTONOZ P,SPIEGELMAN B M.Fat and Beyond:the diverse biology of PPARγ[J].Annu Rev Biochem,2008,77:289-312.
[34]	NIMONKAR A V,WELDON S,GODBOUT K,et al.A lipoprotein lipase-GPI-anchored high-density lipoprotein-binding protein 1 fusion lowers triglycerides in mice:Implications for managing familial chylomicronemia syndrome[J].J Biol Chem,2020, 295(10):2900-2912.
[35]	RUPPERT P M M,KERSTEN S.A lipase fusion feasts on fat[J].J Biol Chem,2020,295(10):2913-2914.
[36]	PÉREZ-TORRES I,GUTIÉRREZ-ALVAREZ Y,GUARNER-LANS V,et al.Intra-abdominal fat adipocyte hypertrophy through a progressive alteration of lipolysis and lipogenesis in metabolic syndrome rats[J].Nutrients,2019,11(7):1529.

miR-106b-5p Regulates the Differentiation of Goat Intramuscular Preadipocytes by Targeting KLF4

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