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1、siRNA对HepG2细胞mcl-1表达的影响530?研究原着?四军医大学(JFourthMilMedUniv)2007,28(6)http://joumal,fmmu.edu.cnsiRNA对HepG2细胞mcl-1表达的影响文章编号:1000-2790(2007)06-05304)4张巍,赵蒲,曹大勇,陶开山,蔡磊,张福琴,窦科峰(第四军医大学:'西京医院肝胆外科,基础部细胞生物学教研室,陕西西安710033)EffectofsiRNAonmcl-1expressioninHepG2cellsZHANGWei,ZHAOPu,CAODa—y0,T
2、AOKai-Shan,CAILei,ZHANGFu—Qin,DOUKe一DepartmentofHepatobiliarySurgery,XijingHospital,Depar-tmentofCellBiology,SchoolofBasicMedicine,FourthMilitaryMedicalUniversity,Xi'an710033,China【Abstract】AIM_,ToinvestigatetheeffectofspecificsiRNAonmcl一1expressioninhepatomacelllineHepG2.METH
3、ODS:HumanhepatomacelllineHepG2wasculturedinvitroandtransfectedwitlltwosiRNAs(differentloci)ofmcl-1bylipo-fectamlne2000.elevelsofmcl-1mRNAexpressionbeforeandaftertransfectionweredetectedbysemi-quantitativereversetran-scriptionpolymerasechainreaction(RT-PCR).einterferingeffectbe
4、tweenthetwosiRNAsandthespecificityandtimeeffectforinterferencewerecompared.ThenthelevelsofMcl一1proteinexpressionbeforeandaftertransfectionweredetectedbyWesternBlot.REsIJIrS:Thelevelofmc/-ImRNAinHepG2cellswasinhibitedbythespecificsiRNAs.eexpressionofracl一1mRNAbegantodecrease24h
5、aftertransfection;andthemostapparentinterferingefficiencywas67.25%48haftertransfection,whichwasmarkedlyhigherthantllatin山ecellstransfectedwitlltllecontrolsiRNAs;at72h,tlleexpressionrestoredalittle.Bo山siRN~(siRNA—FIandsiRNA—F2)fromdifferemlocihadimerfer-ingeffectonmd一1mBNAexpre
6、ssion,andtherewasonlyalittledirencebetween山em.WestemBlotindicatedtllattlleexpres—sionofMcl一1proteinwasdownregulatedinHepG2cellsafterinterferedwitllsiRNA—F..C0lNCLUSII)N:RNAinterferencecandownregulatethelevelofmcl一1mRNAinHepG2cells.AndMcl一1proteinexpressionisalsodownregulatedby
7、specificsiRNA.【Keywords】carcinoma,hepatocellular;mcl一1;RNAinterfer-enCe【摘要】目的:探讨特异性siRNA对肝癌细胞系HepG2中mcl—j表达的影响.方法:将特异性siRNA经脂质体介导转染人肝癌细胞系HepG2;半定量RT—PCR观察干扰前后mcl—J收稿日期:20o6—12-01;接受日期:2006—12-22作者筒介:张巍.硕士生(导师窦科峰).Tel:(029)84775260Email:zhangwd.98@263.netmRNA水平变化,比较对应不同位点的两对siR
8、NA片段对mcl-1的干扰效果,分析RNA干扰的特异性,时效性;WesternBlot检测转染前后Mcl一1蛋白表达情况.
