miR-378 Promoting Lipogenesis and Identification of Target Genes

doi:10.13560/j.cnki.biotech.bull.1985.2020-0848

Abstract

Abstract:

The purpose of this study was to investigate the function of miR-378 in adipocytes and to filter and verify its target genes. 3T3-L1 cells were transfected with miR-378 mimic to validate the function of mir-378 in adipocytes. The potential target genes of miR-378 were identified according to the conservation of target sites and lipid promoting function. The relationship between miR-378 and its targets was verified by microRNA pulldown technology,and then the binding sites of miR-378 and its targets were determined by Dual-Luciferase Reporter Assay System. Some important results were found that miR-378 promoted lipogenesis in adipocytes by increasing lipid synthesis and decreasing lipid decomposition,and the relationship between miR-378 and Runx1t1,Galnt3,RAB10 and the binding sites were determined.

Key words: miR-378, lipogenesis, microRNA pulldown, target genes, target sites

ZHANG Ting-huan, LONG Xi, GUO Zong-yi, CHAI Jie. miR-378 Promoting Lipogenesis and Identification of Target Genes[J]. Biotechnology Bulletin, 2021, 37(2): 80-87.

Figures/Tables 9

References 41

[1]	Bartel DP. MicroRNAs:target recognition and regulatory functions[J]. Cell, 2009,136(2):215-233. doi: 10.1016/j.cell.2009.01.002 URL pmid: 19167326
[2]	Gagan J, Dey BK, Layer R, et al. MicroRNA-378 targets the myogenic repressor MyoR during myoblast differentiation[J]. J Biol Chem, 2011,286(22):19431-19438. URL pmid: 21471220
[3]	Hupkes M, Sotoca AM, Hendriks JM, et al. MicroRNA miR-378 promotes BMP2-induced osteogenic differentiation of mesenchymal progenitor cells[J]. BMC Mol Biol, 2014,15:1. doi: 10.1186/1471-2199-15-1 URL pmid: 24467925
[4]	Eichner LJ, Perry MC, Dufour CR, et al. miR-378* mediates metabolic shift in breast cancer cells via the PGC-1β/ERRγ transcriptional pathway[J]. Cell Metab, 2010,12(4):352-361. doi: 10.1016/j.cmet.2010.09.002 URL pmid: 20889127
[5]	Hou X, Tang Z, Liu H, et al. Discovery of MicroRNAs associated with myogenesis by deep sequencing of serial developmental skeletal muscles in pigs[J]. PLoS One, 2012,7(12):e52123. doi: 10.1371/journal.pone.0052123 URL pmid: 23284895
[6]	Knezevic I, Patel A, Sundaresan NR, et al. A novel cardiomyocyte-enriched microRNA, miR-378, targets insulin-like growth factor 1 receptor:implications in postnatal cardiac remodeling and cell survival[J]. J Biol Chem, 2012,287(16):12913-12926. doi: 10.1074/jbc.M111.331751 URL pmid: 22367207
[7]	Fang J, Song XW, Tian J, et al. Overexpression of microRNA-378 attenuates ischemia-induced apoptosis by inhibiting caspase-3 expression in cardiac myocytes[J]. Apoptosis, 2012,17(4):410-423. doi: 10.1007/s10495-011-0683-0 URL pmid: 22119805
[8]	Liu W, Cao H, Ye C, et al. Hepatic miR-378 targets p110alpha and controls glucose and lipid homeostasis by modulating hepatic insulin signalling[J]. Nat Commun, 2014,5:5684. doi: 10.1038/ncomms6684 URL pmid: 25471065
[9]	Carrer M, Liu N, Grueter CE, et al. Control of mitochondrial metabolism and systemic energy homeostasis by microRNAs 378 and 378*[J]. Proc Natl Acad Sci USA, 2012,109(38):15330-15335. doi: 10.1073/pnas.1207605109 URL pmid: 22949648
[10]	Kahai S, Lee SC, Lee DY, et al. MicroRNA miR-378 regulates nephronectin expression modulating osteoblast differentiation by targeting GalNT-7[J]. PLoS One, 2009,4(10):e7535. doi: 10.1371/journal.pone.0007535 URL pmid: 19844573
[11]	Kim HS, Lee KS, Bae HJ, et al. MicroRNA-31 functions as a tumor suppressor by regulating cell cycle and epithelial-mesenchymal transition regulatory proteins in liver cancer[J]. Oncotarget, 2015,6(10):8089. doi: 10.18632/oncotarget.3512 URL pmid: 25797269
[12]	Zhang C, Liu J, Tan C, et al. microRNA-1827 represses MDM2 to positively regulate tumor suppressor p53 and suppress tumorigenesis[J]. Oncotarget, 2016,7(8):8783. doi: 10.18632/oncotarget.7088 URL pmid: 26840028
[13]	Tan SM, Lieberman J. Capture and identification of miRNA targets by biotin pulldown and RNA-seq[M]//Dassi E(eds)Post-Transcriptional Gene Regulation, New York: Humaoo Press, 2016, 211-228.
[14]	Dragomir MP, Knutsen E, Calin GA. SnapShot:unconventional miRNA functions[J]. Cell, 2018,174(4):1038. doi: 10.1016/j.cell.2018.07.040 URL pmid: 30096304
[15]	Chai J, Chen L, Luo Z, et al. Spontaneous single nucleotide polymorphism in porcine microRNA-378 seed region leads to functional alteration[J]. Biosci Biotechnol Biochem, 2018,82(7):1081-1089. doi: 10.1080/09168451.2018.1459175 URL pmid: 29658390
[16]	汪瑞婷, 宋懿朋, 李常银, 等. miR-378 转基因小鼠脂肪组织的代谢表型分析[J]. 中国细胞生物学学报, 2019,41(7):1377-1386.
	Wang RT, Song YP, Li CY, et al. Metabolomics phenotypes of adipose tissues from miR-378 transgenic mice[J]. Chinese Journal of Cell Biology, 2019,41(7):1377-1386.
[17]	张阳阳, 戴立胜, 周乾, 等. miR-378在牛不同组织中的表达规律及功能分析[J]. 黑龙江动物繁殖, 2014,22(3):15-18.
	Zhang YY, Dai LS, Zhou Q, et al. Expression and functional analysis of mir-378 in different tissues of cattle[J]. Heilongjiang Journal of Animal Reproduction, 2014,22(3):15-18.
[18]	Xu LL, Shi CM, Xu GF, et al. TNF-α, IL-6, and leptin increase the expression of miR-378, an adipogenesis-related microRNA in human adipocytes[J]. Cell Biochemistry and Biophysics, 2014,70(2):771-776. doi: 10.1007/s12013-014-9980-x URL pmid: 24771406
[19]	Gerin I, Bommer GT, Mccoin CS, et al. Roles for miRNA-378/378* in adipocyte gene expression and lipogenesis[J]. Am J Physiol Endocrinol Metab, 2010,299(2):E198-E206. doi: 10.1152/ajpendo.00179.2010 URL pmid: 20484008
[20]	Pan D, Mao C, Quattrochi B, et al. MicroRNA-378 controls classical brown fat expansion to counteract obesity[J]. Nat Commun, 2014,5:4725. doi: 10.1038/ncomms5725 URL pmid: 25145289
[21]	Kim J, Okla M, Erickson A, et al. Eicosapentaenoic acid potentiates brown thermogenesis through FFAR4-dependent up-regulation of miR-30b and miR-378[J]. Journal of Biological Chemistry, 2016,291(39):20551-20562.
[22]	Xu L, Ma X, Verma NK, et al. Ablation of PPAR γ in subcutaneous fat exacerbates age-associated obesity and metabolic decline[J]. Aging Cell, 2018,17(2):e12721.
[23]	刘亚茹, 苗志国, 高明磊, 等. PPARγ 在动物脂肪发育中的研究进展[J]. 黑龙江畜牧兽医, 2019(1):32-35.
	Liu YR, Miao ZG, Gao ML, et al. Research advance on PPARγ in animal adipose tissue[J]. Heilongjiang Animal Science and Veterinary Medicine, 2019(1):32-35.
[24]	Seiri P, Abi A, Soukhtanloo M. PPAR-γ:Its ligand and its regulation by microRNAs[J]. J Cell Biochem, 2019,120(7):10893-10908.
[25]	Schreiber R, Diwoky C, Schoiswohl G, et al. Cold-induced thermogenesis depends on ATGL-mediated lipolysis in cardiac muscle, but not brown adipose tissue[J]. Cell Metab, 2017,26(5):753-763. URL pmid: 28988821
[26]	Schreiber R, Xie H, Schweiger M. Of mice and men:The physiological role of adipose triglyceride lipase(ATGL)[J]. Biochimica et Biophysica Acta(BBA)-Molecular and Cell Biology of Lipids, 2019,1864(6):880-899.
[27]	Sortica D, Rheinheimer J, Moehlecke M, et al. CGI-58 gene expression is decreased in the subcutaneous adipose tissue of patients with obesity[S]. 21st European Congress of Endocrinology:BioScientifica, 2019.
[28]	Korbelius M, Vujic N, Sachdev V, et al. ATGL/CGI-58-dependent hydrolysis of a lipid storage pool in murine enterocytes[J]. Cell Rep, 2019,28(7):1923-1934. doi: 10.1016/j.celrep.2019.07.030 URL pmid: 31412256
[29]	Spiegelman BM. Banting Lecture 2012:Regulation of adipogenesis:toward new therapeutics for metabolic disease[J]. Diabetes, 2013,62(6):1774-1782. doi: 10.2337/db12-1665 URL
[30]	Huang N, Wang J, Xie W, et al. MiR-378a-3p enhances adipogenesis by targeting mitogen-activated protein kinase 1[J]. Biochem Biophys Res Commun, 2015,457(1):37-42. doi: 10.1016/j.bbrc.2014.12.055 URL pmid: 25529446
[31]	Payne VA, Au WS, Lowe CE, et al. C/EBP transcription factors regulate SREBP1c gene expression during adipogenesis[J]. Biochemical Journal, 2010,425(1):215-224.
[32]	Wang SS, Huang HY, Chen SZ, et al. Gdf6 induces commitment of pluripotent mesenchymal C3H10T1/2 cells to the adipocyte lineage[J]. FEBS J, 2013,280(11):2644-2651. doi: 10.1111/febs.12256 URL pmid: 23527555
[33]	Deng K, Ren C, Liu Z, et al. Characterization of RUNX1T1, an adipogenesis regulator in ovine preadipocyte differentiation[J]. Int J Mol Sci, 2018,19(5):1300.
[34]	Sano H, Roach WG, Peck GR, et al. Rab10 in insulin-stimulated GLUT4 translocation[J]. Biochemical Journal, 2008,411(1):89-95.
[35]	Chen Y, Wang Y, Zhang J, et al. Rab10 and myosin-Va mediate insulin-stimulated GLUT4 storage vesicle translocation in adipocytes[J]. J Cell Biol, 2012,198(4):545-560. doi: 10.1083/jcb.201111091 URL pmid: 22908308
[36]	Friesen M, Cowan CA. Adipocyte metabolism and insulin signaling perturbations:insights from genetics[J]. Trends in Endocrinology & Metabolism, 2019,30(6):396-406. doi: 10.1016/j.tem.2019.03.002 URL pmid: 31072658
[37]	Benet-Pag SA, Orlik P, Strom TM, et al. An FGF23 missense mutation causes familial tumoral calcinosis with hyperphosphatemia[J]. Human Molecular Genetics, 2004,14(3):385-390. doi: 10.1093/hmg/ddi034 URL pmid: 15590700
[38]	Ichikawa S, Sorenson AH, Austin AM, et al. Ablation of the Galnt3 gene leads to low-circulating intact fibroblast growth factor 23(Fgf23)concentrations and hyperphosphatemia despite increased Fgf23 expression[J]. Endocrinology, 2009,150(6):2543-2550. doi: 10.1210/en.2008-0877 URL pmid: 19213845
[39]	Awan HM, Shah A, Rashid F, et al. Comparing two approaches of miR-34a target identification, biotinylated-miRNA pulldown vs miRNA overexpression[J]. RNA Biology, 2018,15(1):55-61. doi: 10.1080/15476286.2017.1391441 URL pmid: 29028450
[40]	Phatak P, Donahue JM. Biotinylated micro-RNA pull down assay for identifying miRNA targets[J]. Bio Protocol, 2017,7(9):e2253.
[41]	Li X, Pritykin Y, Concepcion CP, et al. High-resolution in vivo identification of miRNA targets by Halo-Enhanced Ago2 Pulldown[J]. BioRxiv, 2019: 820548.