miR-127-3p对C2C12细胞成肌分化和转录组的影响
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摘要
前期研究表明,miR-127-3p参与骨骼肌细胞分化的调控,但对于miR-127-3p调控的靶基因及其在成肌分化中的作用还不清楚。应用qRT-PCR与细胞形态学方法研究过表达miR-127-3p对C2C12成肌细胞分化及成肌标志基因MyoD、MyoG和Myosin表达的影响;RNA-Seq分析过表达miR-127-3p对成肌细胞转录组的影响,鉴定差异表达基因并对其功能进行初步研究。结果表明,过表达miR-127-3p显著促进C2C12细胞的成肌分化与MyoD、MyoG和Myosin表达;RNA-Seq分析共鉴定到3个差异基因,均为下调基因,其中包括2个转录因子(Irf7和Ddit3)。GO与KEGG分析显示,这些差异基因显著富集于免疫和能量代谢过程/信号通路。生物信息学分析发现,miR-127-3p可与Ddit3基因的CDS区发生碱基互补配对,进而抑制其表达。表明在转录组水平上,miR-127-3p可能通过直接靶向Ddit3基因/转录因子调控免疫与能量代谢类基因的表达,进而调节成肌细胞分化。
MiR-127-3p was involved in myogenic differentiation in our previous study,however its targets and effect on myogenesis were not clear.We examined the effect of miR-127-3p on differentiation of C2C12 myoblasts using qRT-PCR and cell morphology methods.Then we performed a deep sequencing of transcriptome in C2C12 cells to identify the differential expressed(DE)genes and their function involved by bioinformatic analysis.The results demonstrated that overexpression of miR-127-3p promoted myogenic differentiation of C2C12 myoblasts and relative expression of MyoD,MyoG,Myosin.Transcriptome analysis by RNA-Seq illustrated that the C2C12 myoblasts overexpressing miR-127-3p significantly down-regulated expression of 30 genes including two transcription factors(Irf7 and Ddit3),compared with the negative control.Further analysis indicated that these 30 DE genes,which were down-regulated by overexpression of miR-127-3p,were enriched in biological process or pathways involved in immune and energy metabolism.Further analysis indicated that miR-127-3p could target the coding sequence(CDS)of Ddit3,thereby suppress its expression.In conclusion,miR-127-3p could modulate myogenic differentiation of C2C12 myoblasts by regulating expression in target genes associated with immune and energy metabolism at the transcriptome level.MiR-127-3p could inhibit expression of Ddit3 by targeting its CDS.
MiR-127-3p was involved in myogenic differentiation in our previous study,however its targets and effect on myogenesis were not clear.We examined the effect of miR-127-3p on differentiation of C2C12 myoblasts using qRT-PCR and cell morphology methods.Then we performed a deep sequencing of transcriptome in C2C12 cells to identify the differential expressed(DE)genes and their function involved by bioinformatic analysis.The results demonstrated that overexpression of miR-127-3p promoted myogenic differentiation of C2C12 myoblasts and relative expression of MyoD,MyoG,Myosin.Transcriptome analysis by RNA-Seq illustrated that the C2C12 myoblasts overexpressing miR-127-3p significantly down-regulated expression of 30 genes including two transcription factors(Irf7 and Ddit3),compared with the negative control.Further analysis indicated that these 30 DE genes,which were down-regulated by overexpression of miR-127-3p,were enriched in biological process or pathways involved in immune and energy metabolism.Further analysis indicated that miR-127-3p could target the coding sequence(CDS)of Ddit3,thereby suppress its expression.In conclusion,miR-127-3p could modulate myogenic differentiation of C2C12 myoblasts by regulating expression in target genes associated with immune and energy metabolism at the transcriptome level.MiR-127-3p could inhibit expression of Ddit3 by targeting its CDS.
引文
[1]COMAI G,TAJBAKHSH S.Molecular and cellular regulation of skeletal myogenesis[J].Current Topics in Developmental Biology,2014,110:1-73.
[2]LASSAR A B,SKAPEK S X,NOVITCH B.Regulatory mechanisms that coordinate skeletal muscle differentiation and cell cycle withdrawal[J].Current Opinion in Cell Biology,1994,6(6):788-794.
[3]GE Y,CHEN J.MicroRNAs in skeletal myogenesis[J].Cell Cycle,2011,10(3):441-448.
[4]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.
[5]WANG H,WANG B.Extracellular vesicle microRNAs mediate skeletal muscle myogenesis and disease[J].Biomedical Reports,2016,5(3):296-300.
[6]CHEN J F,TAO Y,LI J,et al.MicroRNA-1and micro RNA-206regulate skeletal muscle satellite cell proliferation and differentiation by repressing Pax7[J].Journal of Cell Biology,2010,190(5):867-879.
[7]CHEN J F,MANDEL E M,THOMSON J M,et al.The role of microRNA-1and microRNA-133in skeletal muscle proliferation and differentiation[J].Nature Genetics,2006,38(2):228-233.
[8]DEY B K,GAGAN J DUTTA A.MiR-206and-486induce myoblast differentiation by downregulating Pax7[J].Molecular Cell Biology,2011,31(1):203-214.
[9]NAGUIBNEVA I,AMEYAR-ZAZOUA M,POLESS-KAYA A,et al.The microRNA miR-181targets the homeobox protein Hox-A11during mammalian myoblast differentiation[J].Nature Cell Biology,2006,8(3):278-284.
[10]ZHANG J,YING Z Z,TANG Z L,et al.MicroRNA-148a promotes myogenic differentiation by targeting the ROCK1 gene[J].The Journal of Biology of Chemistry,2012,287(25):21093-21101.
[11]FENG Y,CAO J H,LI X Y,et al.Inhibition of miR-214expression represses proliferation and differentiation of C2C12myoblasts[J].Cell Biochemistry and Function,2011,29(5):378-383.
[12]DEY B K,GAGAN J,YAN Z,et al.MiR-26ais required for skeletal muscle differentiation and regeneration in mice[J].Genes&Development,2012,26(19):2180-2191.
[13]CRIST C G,MONTARRAS D,PALLAFACCHINA G,et al.Muscle stem cell behavior is modified by microRNA-27regulation of Pax3 expression[J].The Proceedings of the National Academy of Sciences of the United States of America,2009,106(32):13383-13387.
[14]GAGAN J,DEY B K,LAYER R,et al.MicroRNA-378targets the myogenic repressor MyoR during myoblast differentiation[J].The Journal of Biology of Chemistry,2011,286(22):19431-19438.
[15]SARKAR S,DEY B K,DUTTA A.MiR-322/424and-503are induced during muscle differentiation and promote cell cycle quiescence and differentiation by down-regulation of Cdc25A[J].Molecular Biology of the Cell,2010,21(13):2138-2149.
[16]GE Y,SUN Y,CHEN J.IGF-Ⅱis regulated by microR-NA-125bin skeletal myogenesis[J].Journal Cell Biology,2011,192(1):69-81.
[17]SEOK H Y,TATSUGUCHI M,CALLIS T E,et al.MiR-155inhibits expression of the MEF2Aprotein to repress skeletal muscle differentiation[J].The Journal of Biology of Chemistry,2011,286(41):35339-35346.
[18]JIA L,LI Y F,WU G F,et al.MiRNA-199a-3p regulates C2C12 myoblast differentiation through IGF-1/AKT/mTOR signal pathway[J].International Journal of Molecular Sciences,2013,15(1):296-308.
[19]ANTONIOU A,MASTROYIANNOPOULOS N P,UN-EY J B,et al.MiR-186inhibits muscle cell differentiation through myogenin regulation[J].The Journal of Biology of Chemestry,2014,289(7):3923-3935.
[20]CARDINALI B,CASTELLANI L,FASANARO P,et al.Microrna-221and microrna-222 modulate differentiation and maturation of skeletal muscle cells[J].PLoS One,2009,4(10):e7607.
[21]CRIPPA S,CASSANO M,MESSINA G,et al.MiR-669a and miR-669q prevent skeletal muscle differentiation in postnatal cardiac progenitors[J].Journal of Cell Biology,2011,193(7):1197-1212.
[22]ZHANG S,ZHAO F,WEI C,et al.Identification and characterization of the miRNA transcriptome of Ovis aries[J].PLoS One,2013,8(3):e58905.
[23]YANG Y L,LI Y,LIANG R Y,et al.Dynamic expression of microRNA-127during porcine prenatal and postnatal skeletal muscle development[J].Journal of Integrative Agriculture,2014,13(6):1331-1339.
[24]LI J,WANG G,JIANG J,et al.Dynamical expression of miR-127-3p in proliferating and differentiating C2C12cells[J].Asian-Australasian Journal of Animal Sciences,2016,29(12):1790-1795.
[25]KIM D,PERTEA G,TRAPNELL C,et al.TopHat2:accurate alignment of transcriptomes in the presence of insertions,deletions and gene fusions[J].Genome Biology,2013,14(4):R36.
[26]TRAPNELL C,ROBERTS A,GOFF L,et al.Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks[J].Nature Protocols,2012,7(3):562-578.
[27]GUTTMAN M,GARBER M,LEVIN J Z,et al.Ab initio reconstruction of cell type-specific transcriptomes in mouse reveals the conserved multi-exonic structure of lincRNAs[J].Nature Biotechnology,2010,28(5):503-510.
[28]TRAPNELL C,WILLIAMS B A,PERTEA G,et al.Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J].Nature Biotechnology,2010,28(5):511-515.
[29]BERKES C A,TAPSCOTT S J.MyoDand the transcriptional control of myogenesis[J].Seminars in Cell&Developmental Biology,2005,16(4):585-595.
[30]GE Y,CHEN J.MicroRNAs in skeletal myogenesis[J].Cell Cycle,2011,10(3):441-448.
[31]SONG G,WANG L.Transcriptional mechanism for the paired miR-433and miR-127genes bynuclear receptors SHP and ERRgamma[J].Nucleic Acids Research,2008,36(18):5727-5735.
[32]BHASKARAN M,WANG Y,ZHANG H,et al.MicroR-NA-127modulates fetal lung development[J].Physiological Genomics,2009,37(3):268-278.
[33]KARPATI G,POULIOT Y,CARPENTER S.Expression of immunoreactive major histocompatibility complex products in human skeletal muscles[J].Annals of Neurology,1988,23(1):64-72.
[34]HONDA H,ROSTAMI A.Expression of major histocompatibility complex classⅠantigens in rat muscle cultures:the possible developmental role in myogenesis[J].The Proceedings of the National Academy of Sciences of the United States of America,1989,86(18):7007-7011.
[35]O’CONNOR R S,STEEDS C M,WISEMAN R W,et al.Phosphocreatine as an energy source for actin cytoskeletal rearrangements during myoblast fusion[J].Journal of Physiology,2008,586(12):2841-2853.
[36]ALTER J,BENGAL E.Stress-induced C/EBP homology protein(CHOP)represses MyoD transcription to delay myoblast differentiation[J].PLoS One,2011,6(12):e29498.
[37]侯金星,雷颖楠,梁玉发,等.chi-miR-4110对奶山羊Smad2基因的靶向调控作用[J].西北农业学报,2018,27(1):10-15.HOU J X,LEI Y N,LIANG Y F,et al.Regulation function of chi-miR-4110on Smad2 gene in dairy goats[J].Acta Agriculturae Boreali-occidentalis Sinica,2018,27(1):10-15.
[38]穆松,钟金城,陈智华等.牛基因组per-microRNA SNP多态性[J].西北农业学报,2011 20,28-33.MU S,ZHONG J CH,CHEN ZH H,et al.Single nucleotide plymorphisms in cattle genomic pre microRNA[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(10):28-33.
[39]DUURSMA A M,KEDDE M,SCHRIER M,et al.MiR-148targets human DNMT3bprotein coding region[J].RNA,2008,14(5):872-877.
[40]ELCHEVA I,GOSWAMI S,NOUBISSI F K,et al.CRD-BP protects the coding region of betaTrCP1mRNA from miR-183-mediated degradation[J].Molecular Cell,2009,35(2):240-246.
[41]YI C,XIE W D,LI F,et al.MiR-143enhances adipogenic differentiation of 3T3-L1cells through targeting the coding region of mouse pleiotrophin[J].FEBS Letters,2011,585(20):3303-3309.
[42]JUNG H M,PATEL R S,PHILLIPS B L,et al.Tumor suppressor miR-375regulates MYC expression via repression of CIP2Acoding sequence through multiple miRNA-mRNA interactions[J].Molecular Biology of the Cell,2013,24(11):1638-1648,S1631-1637.
[43]ZENG Y,PAN Y,LIU H,et al.MiR-20aregulates the PRKG1gene by targeting its coding region in pulmonary arterial smooth muscle cells[J].FEBS Letters,2014,588(24):4677-4685.
[44]SHI L,ZhOU B,LI P,et al.MicroRNA-128targets myostatin at coding domain sequence to regulate myoblasts in skeletal muscle development[J].Cell Signaling,2015,27(9):1895-1904.
[45]QUANN K,JING Y,RIGOUTSOS I.Post-transcriptional regulation of BRCA1through its coding sequence by the miR-15/107group of miRNAs[J].Frontiers in Genetics,2015,6:242.
[46]QIAN J,TU R,YUAN L,et al.Intronic miR-932targets the coding region of its host gene,Drosophila neuroligin2[J].Experimental Cell Research,2016,344(2):183-193.
[47]ZHOU J,DUAN H,XIE Y,et al.MiR-193a-5p targets the coding region of AP-2alpha mRNA and induces cisplatin resistance in bladder cancers[J].Journal of Cancer,2016,7(12):1740-1746.
[48]TONG L,LIN L,WU S,et al.MiR-10a*up-regulates coxsackievirus B3biosynthesis by targeting the 3D-coding sequence[J].Nucleic Acids Research,2013,41(6):3760-3771.
[49]WANG L,QIN Y,TONG L,et al.MiR-342-5p suppresses coxsackievirus B3biosynthesis by targeting the 2C-coding region[J].Antiviral Research,2012,93(2):270-279.
[50]CHEN H,JIANG S,WANG L,et al.Cgi-miR-92dindirectly regulates TNF expression by targeting CDS region of lipopolysaccharide-induced TNF-alpha factor 3(CgLI-TAF3)in oyster Crassostrea gigas[J].Fish Shellfish Immunology,2016,55:577-584.
[2]LASSAR A B,SKAPEK S X,NOVITCH B.Regulatory mechanisms that coordinate skeletal muscle differentiation and cell cycle withdrawal[J].Current Opinion in Cell Biology,1994,6(6):788-794.
[3]GE Y,CHEN J.MicroRNAs in skeletal myogenesis[J].Cell Cycle,2011,10(3):441-448.
[4]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.
[5]WANG H,WANG B.Extracellular vesicle microRNAs mediate skeletal muscle myogenesis and disease[J].Biomedical Reports,2016,5(3):296-300.
[6]CHEN J F,TAO Y,LI J,et al.MicroRNA-1and micro RNA-206regulate skeletal muscle satellite cell proliferation and differentiation by repressing Pax7[J].Journal of Cell Biology,2010,190(5):867-879.
[7]CHEN J F,MANDEL E M,THOMSON J M,et al.The role of microRNA-1and microRNA-133in skeletal muscle proliferation and differentiation[J].Nature Genetics,2006,38(2):228-233.
[8]DEY B K,GAGAN J DUTTA A.MiR-206and-486induce myoblast differentiation by downregulating Pax7[J].Molecular Cell Biology,2011,31(1):203-214.
[9]NAGUIBNEVA I,AMEYAR-ZAZOUA M,POLESS-KAYA A,et al.The microRNA miR-181targets the homeobox protein Hox-A11during mammalian myoblast differentiation[J].Nature Cell Biology,2006,8(3):278-284.
[10]ZHANG J,YING Z Z,TANG Z L,et al.MicroRNA-148a promotes myogenic differentiation by targeting the ROCK1 gene[J].The Journal of Biology of Chemistry,2012,287(25):21093-21101.
[11]FENG Y,CAO J H,LI X Y,et al.Inhibition of miR-214expression represses proliferation and differentiation of C2C12myoblasts[J].Cell Biochemistry and Function,2011,29(5):378-383.
[12]DEY B K,GAGAN J,YAN Z,et al.MiR-26ais required for skeletal muscle differentiation and regeneration in mice[J].Genes&Development,2012,26(19):2180-2191.
[13]CRIST C G,MONTARRAS D,PALLAFACCHINA G,et al.Muscle stem cell behavior is modified by microRNA-27regulation of Pax3 expression[J].The Proceedings of the National Academy of Sciences of the United States of America,2009,106(32):13383-13387.
[14]GAGAN J,DEY B K,LAYER R,et al.MicroRNA-378targets the myogenic repressor MyoR during myoblast differentiation[J].The Journal of Biology of Chemistry,2011,286(22):19431-19438.
[15]SARKAR S,DEY B K,DUTTA A.MiR-322/424and-503are induced during muscle differentiation and promote cell cycle quiescence and differentiation by down-regulation of Cdc25A[J].Molecular Biology of the Cell,2010,21(13):2138-2149.
[16]GE Y,SUN Y,CHEN J.IGF-Ⅱis regulated by microR-NA-125bin skeletal myogenesis[J].Journal Cell Biology,2011,192(1):69-81.
[17]SEOK H Y,TATSUGUCHI M,CALLIS T E,et al.MiR-155inhibits expression of the MEF2Aprotein to repress skeletal muscle differentiation[J].The Journal of Biology of Chemistry,2011,286(41):35339-35346.
[18]JIA L,LI Y F,WU G F,et al.MiRNA-199a-3p regulates C2C12 myoblast differentiation through IGF-1/AKT/mTOR signal pathway[J].International Journal of Molecular Sciences,2013,15(1):296-308.
[19]ANTONIOU A,MASTROYIANNOPOULOS N P,UN-EY J B,et al.MiR-186inhibits muscle cell differentiation through myogenin regulation[J].The Journal of Biology of Chemestry,2014,289(7):3923-3935.
[20]CARDINALI B,CASTELLANI L,FASANARO P,et al.Microrna-221and microrna-222 modulate differentiation and maturation of skeletal muscle cells[J].PLoS One,2009,4(10):e7607.
[21]CRIPPA S,CASSANO M,MESSINA G,et al.MiR-669a and miR-669q prevent skeletal muscle differentiation in postnatal cardiac progenitors[J].Journal of Cell Biology,2011,193(7):1197-1212.
[22]ZHANG S,ZHAO F,WEI C,et al.Identification and characterization of the miRNA transcriptome of Ovis aries[J].PLoS One,2013,8(3):e58905.
[23]YANG Y L,LI Y,LIANG R Y,et al.Dynamic expression of microRNA-127during porcine prenatal and postnatal skeletal muscle development[J].Journal of Integrative Agriculture,2014,13(6):1331-1339.
[24]LI J,WANG G,JIANG J,et al.Dynamical expression of miR-127-3p in proliferating and differentiating C2C12cells[J].Asian-Australasian Journal of Animal Sciences,2016,29(12):1790-1795.
[25]KIM D,PERTEA G,TRAPNELL C,et al.TopHat2:accurate alignment of transcriptomes in the presence of insertions,deletions and gene fusions[J].Genome Biology,2013,14(4):R36.
[26]TRAPNELL C,ROBERTS A,GOFF L,et al.Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks[J].Nature Protocols,2012,7(3):562-578.
[27]GUTTMAN M,GARBER M,LEVIN J Z,et al.Ab initio reconstruction of cell type-specific transcriptomes in mouse reveals the conserved multi-exonic structure of lincRNAs[J].Nature Biotechnology,2010,28(5):503-510.
[28]TRAPNELL C,WILLIAMS B A,PERTEA G,et al.Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J].Nature Biotechnology,2010,28(5):511-515.
[29]BERKES C A,TAPSCOTT S J.MyoDand the transcriptional control of myogenesis[J].Seminars in Cell&Developmental Biology,2005,16(4):585-595.
[30]GE Y,CHEN J.MicroRNAs in skeletal myogenesis[J].Cell Cycle,2011,10(3):441-448.
[31]SONG G,WANG L.Transcriptional mechanism for the paired miR-433and miR-127genes bynuclear receptors SHP and ERRgamma[J].Nucleic Acids Research,2008,36(18):5727-5735.
[32]BHASKARAN M,WANG Y,ZHANG H,et al.MicroR-NA-127modulates fetal lung development[J].Physiological Genomics,2009,37(3):268-278.
[33]KARPATI G,POULIOT Y,CARPENTER S.Expression of immunoreactive major histocompatibility complex products in human skeletal muscles[J].Annals of Neurology,1988,23(1):64-72.
[34]HONDA H,ROSTAMI A.Expression of major histocompatibility complex classⅠantigens in rat muscle cultures:the possible developmental role in myogenesis[J].The Proceedings of the National Academy of Sciences of the United States of America,1989,86(18):7007-7011.
[35]O’CONNOR R S,STEEDS C M,WISEMAN R W,et al.Phosphocreatine as an energy source for actin cytoskeletal rearrangements during myoblast fusion[J].Journal of Physiology,2008,586(12):2841-2853.
[36]ALTER J,BENGAL E.Stress-induced C/EBP homology protein(CHOP)represses MyoD transcription to delay myoblast differentiation[J].PLoS One,2011,6(12):e29498.
[37]侯金星,雷颖楠,梁玉发,等.chi-miR-4110对奶山羊Smad2基因的靶向调控作用[J].西北农业学报,2018,27(1):10-15.HOU J X,LEI Y N,LIANG Y F,et al.Regulation function of chi-miR-4110on Smad2 gene in dairy goats[J].Acta Agriculturae Boreali-occidentalis Sinica,2018,27(1):10-15.
[38]穆松,钟金城,陈智华等.牛基因组per-microRNA SNP多态性[J].西北农业学报,2011 20,28-33.MU S,ZHONG J CH,CHEN ZH H,et al.Single nucleotide plymorphisms in cattle genomic pre microRNA[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(10):28-33.
[39]DUURSMA A M,KEDDE M,SCHRIER M,et al.MiR-148targets human DNMT3bprotein coding region[J].RNA,2008,14(5):872-877.
[40]ELCHEVA I,GOSWAMI S,NOUBISSI F K,et al.CRD-BP protects the coding region of betaTrCP1mRNA from miR-183-mediated degradation[J].Molecular Cell,2009,35(2):240-246.
[41]YI C,XIE W D,LI F,et al.MiR-143enhances adipogenic differentiation of 3T3-L1cells through targeting the coding region of mouse pleiotrophin[J].FEBS Letters,2011,585(20):3303-3309.
[42]JUNG H M,PATEL R S,PHILLIPS B L,et al.Tumor suppressor miR-375regulates MYC expression via repression of CIP2Acoding sequence through multiple miRNA-mRNA interactions[J].Molecular Biology of the Cell,2013,24(11):1638-1648,S1631-1637.
[43]ZENG Y,PAN Y,LIU H,et al.MiR-20aregulates the PRKG1gene by targeting its coding region in pulmonary arterial smooth muscle cells[J].FEBS Letters,2014,588(24):4677-4685.
[44]SHI L,ZhOU B,LI P,et al.MicroRNA-128targets myostatin at coding domain sequence to regulate myoblasts in skeletal muscle development[J].Cell Signaling,2015,27(9):1895-1904.
[45]QUANN K,JING Y,RIGOUTSOS I.Post-transcriptional regulation of BRCA1through its coding sequence by the miR-15/107group of miRNAs[J].Frontiers in Genetics,2015,6:242.
[46]QIAN J,TU R,YUAN L,et al.Intronic miR-932targets the coding region of its host gene,Drosophila neuroligin2[J].Experimental Cell Research,2016,344(2):183-193.
[47]ZHOU J,DUAN H,XIE Y,et al.MiR-193a-5p targets the coding region of AP-2alpha mRNA and induces cisplatin resistance in bladder cancers[J].Journal of Cancer,2016,7(12):1740-1746.
[48]TONG L,LIN L,WU S,et al.MiR-10a*up-regulates coxsackievirus B3biosynthesis by targeting the 3D-coding sequence[J].Nucleic Acids Research,2013,41(6):3760-3771.
[49]WANG L,QIN Y,TONG L,et al.MiR-342-5p suppresses coxsackievirus B3biosynthesis by targeting the 2C-coding region[J].Antiviral Research,2012,93(2):270-279.
[50]CHEN H,JIANG S,WANG L,et al.Cgi-miR-92dindirectly regulates TNF expression by targeting CDS region of lipopolysaccharide-induced TNF-alpha factor 3(CgLI-TAF3)in oyster Crassostrea gigas[J].Fish Shellfish Immunology,2016,55:577-584.