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1、[键入文字]MiR-155通过SOCS1抑制HBV蛋白的表达蔡启茵1,2，任广立1，张卫云1，冯宇鹏1，马恒灏1（510010广州,广州军区广州总医院儿科1;510405广州,广州中医药大学2）[摘要]目的观察miR-155对SOCS1mRNA及蛋白表达水平的影响，探究miR-155对HBsAg、HBeAg表达的抑制作用。方法从HepG2.2.15细胞中提取基因组，经PCR扩增得到miR-155前体序列，纯化并连接入pmR-mCherry载体。重构载体pmiR-155经去内毒素提取后，转染到HepG2.2.15细胞，分别设立重
2、组组（pmiR-155质粒)，空载组（pmR-mCherry质粒）、转染试剂组、空白组。应用荧光实时定量PCR检测各组miR-155的表达，RT-PCR检测各组SOCS1mRNA的变化，Westernblot检测各组SOCS1蛋白的变化，ELISA检测各组HBsAg、HBeAg表达变化。结果荧光实时定量PCR结果示：以空白组细胞内表达的miR-155量为基准，重组组miR-155表达量（519.43±52.10）明显高于空载组（24.24±16.7）及转染试剂组（35.04±26.09）（P＜0.05）；RT-PCR结果示：重
3、组组SOCS1mRNA的表达量（0.63±0.91）显著低于空白组（P＜0.05）；Westernblot结果示：重组组内SOCS1蛋白表达量显著低于空白组；以空白组的表达量为基准，重组组对HBsAg和HBeAg蛋白表达的抑制率分别为（55.62±3.77）%、（47.87±2.46）%（P＜0.01）。结论过表达miR-1555可能通过可能通过调控SOCS1的表达，从而抑制HBsAg、HBeAg的表达。[关键词]microRNA-155；细胞因子信号转导抑制因子1；乙型肝炎表面抗原；乙型肝炎e抗原[中图法分类号]R512.6
4、+2[文献标志码]AmiR-155functionsasananti-virusmicroRNAinHepatitisBvirusbytargetingSOCS1CaiQiyin,RenGuangli,ZhangWeiyun,FengYupeng,MaHenghao(1DepartmentofPediatrics,GeneralHospitalofGuangzhouMilitaryCommand,Guangzhou,Guangdong,510010,China;2GuangzhouuniversityofChinesemedi
5、cine，Guangzhou,Guangdong,510405,China)[Abstract]ObjectiveToinvestigatethemechanismsofmiR-155regulatingSOCS1intheinhibitionofHBsAgandHBeAg.MethodsThepre-miR-155wasamplifiedfromtotalDNAofHepG2.2.15byPCR.ThetargetgenefragmentwasdigestedandclonedintothepmR-mCherryplasmi
6、d.PmiR-155wastransfectedintoHepG2.2.15cellsbyliposome-mediatedmethod.Theemptyplasmid,thereagentgroupanduntreatedcellsweresetascontrols.Firstofall,theexpressionofmiR-155wasdetectedbythereal-timequantitivePCR.Secondly,theexpressionofSOCS1mRNAweredetectedbyRT-PCR,andth
7、entheexpressionofSOCS1proteinwereinvestigatedbyWesternblot.Finally,theexpressionofHBsAgandHBeAgweredetectedbyELISA.ResultsThepmiR-155eukaryoticover-expressionvectorwassuccessfullyconstructed.MiR-155levelofHepG2.2.15cellswhichweretransfectedwiththerecombinantplasmid（
8、519.43±52.10）（P＜0.05）wasobviouslyhigherthancontrols.RT-PCRshowedtheexpressionofSOCS1mRNAwerelessthanothersgroups.TheexpressionofSOCS1prote
