慢病毒介导shRNA干扰MAT2A与MAT2B基因抑制猪肌内脂肪细胞分化
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摘要
【目的】腺苷甲硫氨酸转移酶(MAT)在ATP的作用下,催化生成体内重要的甲基供体S-腺苷甲硫氨酸(SAMe)。通过构建慢病毒介导的pLenti-H1干扰载体,探究腺苷甲硫氨酸转移酶MAT2A和MAT2B对猪肌内脂肪细胞分化的影响。【方法】无菌条件下采集3—7日龄小猪背最长肌,采用差速贴壁法分离猪肌内脂肪细胞。根据GenBank中猪MAT2A基因序列(AccessionNo.NM_001167650.1)和MAT2B基因序列(AccessionNo.NM_001142832.1),获得其CDS序列。利用Invitrogen公司在线软件BLOCK-iTTM RNAi Designer分别设计shRNA靶序列,将合成的单链寡核苷酸退火形成双链,与经过Bam H I和Xho I (TaKaRa)双酶切后的pLenti-Hl载体连接,转化,并提取质粒进行酶切和测序鉴定。采用X-tremeGENE-HP DNA转染试剂与测序成功的重组质粒以及包装质粒(CMV-Δ8.9和CMV-VSVG)共转染293T细胞,48 h后观察绿色荧光蛋白(GFP)的表达,并进行滴度测定。在猪原代脂肪细胞密度达到70%—80%时,侵染病毒;细胞密度融合时诱导分化。提取分化第8天的猪肌内脂肪细胞的RNA,按照反转录试剂盒操作说明进行反转录,合成cDNA第一链。采用primer primer 5软件设计MAT2A、MAT2B、PPARγ、aP2、CEBP/α、β-actin基因的定量引物,实时定量和Western blot试验检测MAT2A和MAT2B基因的干扰效率。油红O染色和实时定量鉴定MAT2A和MAT2B基因对猪肌内脂肪脂质积累的影响。【结果】酶切及测序证明重组慢病毒载体pLenti-Hl-MAT2A/MAT2B构建成功;包装的慢病毒sh-MAT2A和sh-MAT2B病毒滴度分别为6.7×107和7×107 pfu/mL,侵染肌内脂肪细胞72 h后,可出现90%的绿色荧光蛋白(GFP),表明所包装的病毒可满足侵染猪前体肌内脂肪细胞需要。实时定量结果显示其显著抑制了MAT2A和MAT2B的m RNA水平表达,其干扰效率分别在70%和60%以上;进一步采用Western blot试验及蛋白分析表明,干扰MAT2A基因后,MAT2A蛋白表达水平降低40%左右,差异达到极显著水平(P<0.01);干扰MAT2B基因后,MAT2B蛋白表达水平降低25%左右,差异达到显著水平(P<0.05)。油红O染色和吸光度定量结果显示,与sh-scramble对照组相比,干扰MAT2A和MAT2B基因后,脂滴聚积能力显著抑制猪肌内脂肪细胞脂质积累。实时定量结果显示,干扰MAT2A和MAT2B基因抑制成脂标志基因PPARγ,aP 2及CEBP/α的表达。【结论】成功构建了猪MAT2A和MAT2B基因的慢病毒sh-MAT2A和sh-MAT2B干扰载体。获得的重组慢病毒感染细胞后,能有效降低猪肌内前体脂肪细胞内MAT2A和MAT2B基因的m RNA和蛋白水平表达;进一步试验证明,干扰MAT2A及MAT2B基因后,显著抑制猪肌内脂肪细胞的脂质积累以及成脂关键基因表达。
【Objective】 Methionine adenosyltransferase(MAT), an essential cellular enzyme responsible for S-adenosylmethionine biosynthesis. The purpose of this study was to construct the lentivirus-mediated pLenti-H1 vector and investigated the effect of adenosine methionine transferase MAT2 A and MAT2 B on the differentiation of porcine intramuscular adipocytes.【Method】Porcine longissimus dorsi tissue was collected from 3 to 7 days piglets under sterile conditions and isolated the porcine intramuscular preadipocytes by differential adhesion method. According to MAT2 A gene(Accession No.NM_001167650.1) and MAT2 B gene sequence(Accession No.NM_001142832.1), Invitrogen online software BLOCK-iTTM RNAi Designer was used to respectively design MAT2 A and MAT2 B shRNA target sequence. The synthesized single-chain oligonucleotide was annealed to form double strands DNA, and was ligated with pLenti-Hl vector after Bam H I and Xho I(TaKaRa) double digestion, then the plasmid DNA was extracted and further identified by restriction enzyme digestion and sequencing. X-tremeGENE-HP DNA transfection reagent was used to co-transfect 293 T cells with the correct recombinant plasmid and packaging plasmid(CMV-Δ8.9 and CMV-VSVG). The expression of green fluorescent protein(GFP) was observed after 48 h and then the virus titer was tested. The primary porcine intramuscular preadipocytes were infected with viruses at 70% to 80% cell density and induced adipogenic differentiation when cells at full confluence. Total RNA was extracted from adipocytes and the mRNA reverse transcription was performed by using a Revert Aid First-strand c DNA Synthesis Kit. The gene primer sequences of MAT2 A, MAT2 B, PPARγ, aP2, CEBP/α and β-actin were designed by Primer primer 5.The interference efficiency of MAT2 A and MAT2 B gene on the porcine intramuscular preadipocytes was detected by Real-time quantitative PCR and Western blot. The effects of MAT2 A and MAT2 B on porcine intramuscular adipogenesis were detected by Oil O stating and RT-qPCR. 【Result】Recombinant lentivirus vector pLenti-Hl-MAT2 A/MAT2 B was successfully constructed. The virus titer of lentivirus sh-MAT2 A and sh-MAT2 B were respectively 6.7×107 pfu/m L and 7×107 pfu/mL. After preadipocytes infected with adenovirus for 72 h, Microscopic fluorescence imaging showed 90% of green fluorescent protein(GFP)appeared, indicated that porcine intramuscular preadipocytes were successfully infected by lentivirus sh-MAT2 A and sh-MAT2 B.Real-time qPCR results showed that the expression of MAT2 A and MAT2 B was separately decreased 70% and 60% compared with sh-scramble. The Western blot experiment and protein analysis showed that interference with MAT2 A, the expression of MAT2 A had an approximately 40% decrease and the difference reached high significant level(P<0.01). The expression of MAT2 B had a 25%drop when inhibited with MAT2 B and the different tendency towards statistical significance(P<0.05). Oil O staining and quantitative analysis showed that, the lipid accumulation of porcine intramuscular adipocytes markedly reduced upon silencing of MAT2 A and MAT2 B. Real-time qPCR results showed that knockdown of MAT2 A and MAT2 B inhibited the mRNA levels of PPARγ,aP2 and CEBP/α.【Conclusion】 Lentivirus-mediate sh-MAT2 A and sh-MAT2 B interference system was successfully esablished. The recombinant lentivirus can effectively reduce the m RNA and protein expression of MAT2 A and MAT2 B in porcine intramuscular adipocytes. Further experiments showed that interfering with MAT2 A and MAT2 B genes, the lipid accumulation and adipogenic key genes expression in porcine intramuscular adipocytes were significantly inhibited.
【Objective】 Methionine adenosyltransferase(MAT), an essential cellular enzyme responsible for S-adenosylmethionine biosynthesis. The purpose of this study was to construct the lentivirus-mediated pLenti-H1 vector and investigated the effect of adenosine methionine transferase MAT2 A and MAT2 B on the differentiation of porcine intramuscular adipocytes.【Method】Porcine longissimus dorsi tissue was collected from 3 to 7 days piglets under sterile conditions and isolated the porcine intramuscular preadipocytes by differential adhesion method. According to MAT2 A gene(Accession No.NM_001167650.1) and MAT2 B gene sequence(Accession No.NM_001142832.1), Invitrogen online software BLOCK-iTTM RNAi Designer was used to respectively design MAT2 A and MAT2 B shRNA target sequence. The synthesized single-chain oligonucleotide was annealed to form double strands DNA, and was ligated with pLenti-Hl vector after Bam H I and Xho I(TaKaRa) double digestion, then the plasmid DNA was extracted and further identified by restriction enzyme digestion and sequencing. X-tremeGENE-HP DNA transfection reagent was used to co-transfect 293 T cells with the correct recombinant plasmid and packaging plasmid(CMV-Δ8.9 and CMV-VSVG). The expression of green fluorescent protein(GFP) was observed after 48 h and then the virus titer was tested. The primary porcine intramuscular preadipocytes were infected with viruses at 70% to 80% cell density and induced adipogenic differentiation when cells at full confluence. Total RNA was extracted from adipocytes and the mRNA reverse transcription was performed by using a Revert Aid First-strand c DNA Synthesis Kit. The gene primer sequences of MAT2 A, MAT2 B, PPARγ, aP2, CEBP/α and β-actin were designed by Primer primer 5.The interference efficiency of MAT2 A and MAT2 B gene on the porcine intramuscular preadipocytes was detected by Real-time quantitative PCR and Western blot. The effects of MAT2 A and MAT2 B on porcine intramuscular adipogenesis were detected by Oil O stating and RT-qPCR. 【Result】Recombinant lentivirus vector pLenti-Hl-MAT2 A/MAT2 B was successfully constructed. The virus titer of lentivirus sh-MAT2 A and sh-MAT2 B were respectively 6.7×107 pfu/m L and 7×107 pfu/mL. After preadipocytes infected with adenovirus for 72 h, Microscopic fluorescence imaging showed 90% of green fluorescent protein(GFP)appeared, indicated that porcine intramuscular preadipocytes were successfully infected by lentivirus sh-MAT2 A and sh-MAT2 B.Real-time qPCR results showed that the expression of MAT2 A and MAT2 B was separately decreased 70% and 60% compared with sh-scramble. The Western blot experiment and protein analysis showed that interference with MAT2 A, the expression of MAT2 A had an approximately 40% decrease and the difference reached high significant level(P<0.01). The expression of MAT2 B had a 25%drop when inhibited with MAT2 B and the different tendency towards statistical significance(P<0.05). Oil O staining and quantitative analysis showed that, the lipid accumulation of porcine intramuscular adipocytes markedly reduced upon silencing of MAT2 A and MAT2 B. Real-time qPCR results showed that knockdown of MAT2 A and MAT2 B inhibited the mRNA levels of PPARγ,aP2 and CEBP/α.【Conclusion】 Lentivirus-mediate sh-MAT2 A and sh-MAT2 B interference system was successfully esablished. The recombinant lentivirus can effectively reduce the m RNA and protein expression of MAT2 A and MAT2 B in porcine intramuscular adipocytes. Further experiments showed that interfering with MAT2 A and MAT2 B genes, the lipid accumulation and adipogenic key genes expression in porcine intramuscular adipocytes were significantly inhibited.
引文
[1]BERNS K,HIJMANS E M,MULLENDERS J,BRUMMELKAMP TR,VELDS A,HEIMERIKX M,KERKHOVEN R M,MADIREDJOM,NIJKAMP W,WEIGELT B,AGAMI R,GE W,CAVET G,LINSLEY P S,BEIJERSBERGEN R L,BERNARDS R.A large-scale RNAi screen in human cells identifies new components of the p53pathway.Nature,2004,428(6981):431-437.
[2]LO-BIANCO C,SCHNEIDER B L,BAUER M,SAJADI A,BRICEA,IWATSUBO T,AEBISCHER P.Lentiviral vector delivery of parkin prevents dopaminergic degeneration in an alpha-synuclein rat model of Parkinson's disease.Proceedings of the National Academy of Sciences of the United States of America,2004,101(50):17510-17515.
[3]AMARZGUIOUI M,ROSSI J,KIM D.Approaches for chemically synthesized siRNA and vector-mediated RNAi.FEBS Lett,2005,579(26):5974-5981.
[4]PADDISON P J,CAUDY A A,BERNSTEIN E,HANNON G J,CONKLIN D S.Short hairpin RNAs(shRNAs)induce sequencespecific silencing in mammalian cells.Genes Development,2002,16(8):948-958.
[5]GARCIA-TREVIJANO E R,LATASA M U,CARRETERO M V,BERASAIN C,MATO J M,AVILA M A.S-adenosylmethionine regulates MAT1A and MAT2A gene expression in cultured rathepatocytes:a new role for S-adenosylmethionine in the maintenance of the differentiated status of the liver.FASEB Journal,2000,14(15):2511-2518.
[6]DE LA ROSA J,OSTROWSKI J,HRYNIEWICZ M.Chromosomal localization and catalytic properties of the recombinant alpha subunit of human lymphocyte methionine adenosyltransferase.Journal of Biological Chemistry,1995,270(37):21860-21868.
[7]MATO J M,LU S C.Role of S-adenosyl-L-methionine in liver health and injury.Hepatology,2007,45(5):1306-1312.
[8]YANG H,HUANG Z Z.WANG J,LU S C.The role of c-Myb and Sp1 in the up-regulation of methionine adenosyltransferase 2A gene expression in human hepatocellular carcinoma.FASEB Journal,2001,15(9):1507-1516.
[9]YANG H,SADDA M R,YU V,ZENG Y,LEE T D,OU X,CHENL,LU S C.Induction of human methionine adenosyltransferase2A expression by tumor necrosis factor alpha.Role of NF-kappa B and AP-1.Journal of Biological Chemistry,2003,278(51):50887-50896.
[10]KATOH Y,IKURA T,HOSHIKAWA Y,TASHIRO S,ITO T,OHTAM,KERA Y,NODA T,IGARASHI K.Methionine adenosyltransferase II serves as a transcriptional corepressor of Maf oncoprotein.Molecular Cell,2011,41(5):554-566.
[11]TOMASI M L,RYOO M,RAMANI K,TOMASI I,GIORDANO P,MATO J M,LU S C.Methionine adenosyltransferaseα2 sumoylation positively regulate Bcl-2 expression in human colon and liver cancer cells.Oncotarget,2015,6(35):37706-37723.
[12]RAMANI K,TOMASI M L.Transcriptional regulation of methionine adenosyltransferase 2A by peroxisome proliferator-activated receptors in rat hepatic stellate cells.Hepatology,2012,55(6):1942-1953.
[13]KERA Y,KATOH Y,OHTA M,MATSUMOTO M,TAKANO-YAMAMOTO T,IGARASHI K.Methionine adenosyltransferase II-dependent histone H3K9 methylation at the COX-2 gene locus.Journal of Biological Chemistry,2013,288(19):13592-13601.
[14]LIU L,YIN J,LI W,LIU K,PENG Y,TAN P,MA R Z.Construction of a bacterial artificial chromosome library for the Rongchang pig breed and its use for the identification of genes involved in intramuscular fat deposition.Biochemical and Biophysical Research Communications,2010,391(2):1280-1284.
[15]FANG Q,YIN J,LI F,ZHANG J,WATFORD M.Characterization of methionine adenosyltransferase 2βgene expression in skeletal muscle and subcutaneous adipose tissue from obese and lean pigs.Molecular Biology Reports,2010,37(5):2517-2524.
[16]KRUEGER U,BERGAUER T,KAUFMANN B,WOLTER I,PILKS,HEIDER-FABIAN M,KIRCH S,ARTZ-OPPITZ C,ISSELHORSTM,KONRAD J.Insights into effective RNAi gained from large-scale siRNA validation screening.Oligonucleotides,2007,17(2):237-250.
[17]PFEIFER A,HOFMANN A.Lentiviral transgenesis.Methods Molecular Biology,2009,530(3):391-405.
[18]POESCHLA E,CORBEAU P,WONGSTAAL F.Development of HIV vectors for antiHIV gene therapy.Proceedings of the National Academy of Sciences of the United States of America,1996,93(21):11395-11399.
[19]CHENG J,SONG Z Y,PU L,YANG H,ZHENG J M,ZHANG Z Y,SHI X E,YANG G S.Retinol binding protein 4 affects the adipogenesis of porcine preadipocytes through insulin signaling pathways.Biochemistry and Cell Biology,2013,91(4):236-243.
[20]YANG H,CHENG J,SONG Z,LI X,ZHANG Z,MAI Y,PANG W,SHI X,YANG G.The anti-adipogenic effect of PGRN on porcine preadipocytes involves ERK1,2 mediated PPARγphosphorylation.Molecular Biology Reports,2013,40(12):6863-6872.
[21]FORTIN A,ROBERTSON W M,TONG K W.The eating quality of Canadian pork and its relationship with intramuscular fat.Meat Sciences,2005,69(2):297-305.
[22]赵存真,李黄琨,马云,杨公社.腺病毒介导的MAT2B过表达促进猪肌内脂肪细胞分化.畜牧兽医学报,2017,48(5):802-809.ZHAO C Z,LI H K,MA Y,YANG G S.MAT2B overexpression mediated by adenovirus promoting porcine intramuscular preadipocyte differentiation.Acta Veterinaria et Zootechnica Sinica,2017,48(5):802-809.(in Chinese)
[23]ZHAO C Z,CHEN X C,WU W J,WANG W S,PANG W J,YANG GS.MAT2B promotes adipogenesis by modulating SAMe levels and activating AKT/ERK pathway during porcine intramuscular preadipocyte differentiation.Experimental Cell Research,2016,344(1):11-21.
[24]DE LA ROSA J,OSTROWSKI J,HRYNIEWICZ M M,KREDICH NM,KOTB M,LEGROS H L J R,VALENTINE M,GELLER A M.Chromosomal localization and catalytic properties of the recombinant alpha subunit of human lymphocyte methionine adenosyltransferase.Journal of Biological Chemistry,1995,270(37):21860-21868.
[25]YANG H,SADDA M R,YU V,ZENG Y,LEE T D,OU X,CHENL,LU S C.Induction of human methionine adenosyltransferase2A expression by tumor necrosis factor alpha.Role of NF-kappa B and AP-1.Journal of Biological Chemistry,2003,278(51):50887-50896.
[26]TOMASI M L,RYOO M,RAMANI K,TOMASI I,GIORDANO P,MATO J M,LU S C.Methionine adenosyltransferaseα2 sumoylation positively regulate Bcl-2 expression in human colon and liver cancer cells.Oncotarget,2015,6(35):37706-37723.
[27]RAMANI K,LU S C.Methionine adenosyltransferases in liver health and diseases.Liver Research,2017,1(2):103-111.
[28]BOTTIGLIERI T.S-Adenosyl-L-methionine(SAMe):from the bench to the bedside--molecular basis of a pleiotrophic molecule.American Journal of Clinical Nutrition,2002,76(5):1151S-7S.
[29]MATO J M,LU S C.Role of S-adenosyl-L-methionine in liver health and injury.Hepatology,2007,45(5):1306-1312.
[30]MARTA V R,MARTA I L,JUAN J L,ANA M A,MARC T,AGUSTíN F F,JOSéL L,DAVID M A,MARíA B,MARC T,ZIGMUND L,CONRAD W,SHELLY C L,MANEL E,RHONA M,KRISTJáN R J,MARIO F F,JOSéM M,ASHWIN W.S-adenosylmethionine levels regulate the schwann cell DNA methylome.Neuron,2014,81(5):1024-1039.
[2]LO-BIANCO C,SCHNEIDER B L,BAUER M,SAJADI A,BRICEA,IWATSUBO T,AEBISCHER P.Lentiviral vector delivery of parkin prevents dopaminergic degeneration in an alpha-synuclein rat model of Parkinson's disease.Proceedings of the National Academy of Sciences of the United States of America,2004,101(50):17510-17515.
[3]AMARZGUIOUI M,ROSSI J,KIM D.Approaches for chemically synthesized siRNA and vector-mediated RNAi.FEBS Lett,2005,579(26):5974-5981.
[4]PADDISON P J,CAUDY A A,BERNSTEIN E,HANNON G J,CONKLIN D S.Short hairpin RNAs(shRNAs)induce sequencespecific silencing in mammalian cells.Genes Development,2002,16(8):948-958.
[5]GARCIA-TREVIJANO E R,LATASA M U,CARRETERO M V,BERASAIN C,MATO J M,AVILA M A.S-adenosylmethionine regulates MAT1A and MAT2A gene expression in cultured rathepatocytes:a new role for S-adenosylmethionine in the maintenance of the differentiated status of the liver.FASEB Journal,2000,14(15):2511-2518.
[6]DE LA ROSA J,OSTROWSKI J,HRYNIEWICZ M.Chromosomal localization and catalytic properties of the recombinant alpha subunit of human lymphocyte methionine adenosyltransferase.Journal of Biological Chemistry,1995,270(37):21860-21868.
[7]MATO J M,LU S C.Role of S-adenosyl-L-methionine in liver health and injury.Hepatology,2007,45(5):1306-1312.
[8]YANG H,HUANG Z Z.WANG J,LU S C.The role of c-Myb and Sp1 in the up-regulation of methionine adenosyltransferase 2A gene expression in human hepatocellular carcinoma.FASEB Journal,2001,15(9):1507-1516.
[9]YANG H,SADDA M R,YU V,ZENG Y,LEE T D,OU X,CHENL,LU S C.Induction of human methionine adenosyltransferase2A expression by tumor necrosis factor alpha.Role of NF-kappa B and AP-1.Journal of Biological Chemistry,2003,278(51):50887-50896.
[10]KATOH Y,IKURA T,HOSHIKAWA Y,TASHIRO S,ITO T,OHTAM,KERA Y,NODA T,IGARASHI K.Methionine adenosyltransferase II serves as a transcriptional corepressor of Maf oncoprotein.Molecular Cell,2011,41(5):554-566.
[11]TOMASI M L,RYOO M,RAMANI K,TOMASI I,GIORDANO P,MATO J M,LU S C.Methionine adenosyltransferaseα2 sumoylation positively regulate Bcl-2 expression in human colon and liver cancer cells.Oncotarget,2015,6(35):37706-37723.
[12]RAMANI K,TOMASI M L.Transcriptional regulation of methionine adenosyltransferase 2A by peroxisome proliferator-activated receptors in rat hepatic stellate cells.Hepatology,2012,55(6):1942-1953.
[13]KERA Y,KATOH Y,OHTA M,MATSUMOTO M,TAKANO-YAMAMOTO T,IGARASHI K.Methionine adenosyltransferase II-dependent histone H3K9 methylation at the COX-2 gene locus.Journal of Biological Chemistry,2013,288(19):13592-13601.
[14]LIU L,YIN J,LI W,LIU K,PENG Y,TAN P,MA R Z.Construction of a bacterial artificial chromosome library for the Rongchang pig breed and its use for the identification of genes involved in intramuscular fat deposition.Biochemical and Biophysical Research Communications,2010,391(2):1280-1284.
[15]FANG Q,YIN J,LI F,ZHANG J,WATFORD M.Characterization of methionine adenosyltransferase 2βgene expression in skeletal muscle and subcutaneous adipose tissue from obese and lean pigs.Molecular Biology Reports,2010,37(5):2517-2524.
[16]KRUEGER U,BERGAUER T,KAUFMANN B,WOLTER I,PILKS,HEIDER-FABIAN M,KIRCH S,ARTZ-OPPITZ C,ISSELHORSTM,KONRAD J.Insights into effective RNAi gained from large-scale siRNA validation screening.Oligonucleotides,2007,17(2):237-250.
[17]PFEIFER A,HOFMANN A.Lentiviral transgenesis.Methods Molecular Biology,2009,530(3):391-405.
[18]POESCHLA E,CORBEAU P,WONGSTAAL F.Development of HIV vectors for antiHIV gene therapy.Proceedings of the National Academy of Sciences of the United States of America,1996,93(21):11395-11399.
[19]CHENG J,SONG Z Y,PU L,YANG H,ZHENG J M,ZHANG Z Y,SHI X E,YANG G S.Retinol binding protein 4 affects the adipogenesis of porcine preadipocytes through insulin signaling pathways.Biochemistry and Cell Biology,2013,91(4):236-243.
[20]YANG H,CHENG J,SONG Z,LI X,ZHANG Z,MAI Y,PANG W,SHI X,YANG G.The anti-adipogenic effect of PGRN on porcine preadipocytes involves ERK1,2 mediated PPARγphosphorylation.Molecular Biology Reports,2013,40(12):6863-6872.
[21]FORTIN A,ROBERTSON W M,TONG K W.The eating quality of Canadian pork and its relationship with intramuscular fat.Meat Sciences,2005,69(2):297-305.
[22]赵存真,李黄琨,马云,杨公社.腺病毒介导的MAT2B过表达促进猪肌内脂肪细胞分化.畜牧兽医学报,2017,48(5):802-809.ZHAO C Z,LI H K,MA Y,YANG G S.MAT2B overexpression mediated by adenovirus promoting porcine intramuscular preadipocyte differentiation.Acta Veterinaria et Zootechnica Sinica,2017,48(5):802-809.(in Chinese)
[23]ZHAO C Z,CHEN X C,WU W J,WANG W S,PANG W J,YANG GS.MAT2B promotes adipogenesis by modulating SAMe levels and activating AKT/ERK pathway during porcine intramuscular preadipocyte differentiation.Experimental Cell Research,2016,344(1):11-21.
[24]DE LA ROSA J,OSTROWSKI J,HRYNIEWICZ M M,KREDICH NM,KOTB M,LEGROS H L J R,VALENTINE M,GELLER A M.Chromosomal localization and catalytic properties of the recombinant alpha subunit of human lymphocyte methionine adenosyltransferase.Journal of Biological Chemistry,1995,270(37):21860-21868.
[25]YANG H,SADDA M R,YU V,ZENG Y,LEE T D,OU X,CHENL,LU S C.Induction of human methionine adenosyltransferase2A expression by tumor necrosis factor alpha.Role of NF-kappa B and AP-1.Journal of Biological Chemistry,2003,278(51):50887-50896.
[26]TOMASI M L,RYOO M,RAMANI K,TOMASI I,GIORDANO P,MATO J M,LU S C.Methionine adenosyltransferaseα2 sumoylation positively regulate Bcl-2 expression in human colon and liver cancer cells.Oncotarget,2015,6(35):37706-37723.
[27]RAMANI K,LU S C.Methionine adenosyltransferases in liver health and diseases.Liver Research,2017,1(2):103-111.
[28]BOTTIGLIERI T.S-Adenosyl-L-methionine(SAMe):from the bench to the bedside--molecular basis of a pleiotrophic molecule.American Journal of Clinical Nutrition,2002,76(5):1151S-7S.
[29]MATO J M,LU S C.Role of S-adenosyl-L-methionine in liver health and injury.Hepatology,2007,45(5):1306-1312.
[30]MARTA V R,MARTA I L,JUAN J L,ANA M A,MARC T,AGUSTíN F F,JOSéL L,DAVID M A,MARíA B,MARC T,ZIGMUND L,CONRAD W,SHELLY C L,MANEL E,RHONA M,KRISTJáN R J,MARIO F F,JOSéM M,ASHWIN W.S-adenosylmethionine levels regulate the schwann cell DNA methylome.Neuron,2014,81(5):1024-1039.