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1、siRNA抑制Ku80表达后对宫颈癌HeLa细胞增殖的影响252《癌症》ChineseJournalofCancer,2007,26(3):252—257siRNA抑制Ku80表达后对宫颈癌HeLa细胞增殖的影响庄亮,于世英,黄晓园,高庆蕾,熊EffectofKu80ExpressionInhibitionbyProliferationofCervicalCarcinomaCellZHUANGLiang,YUShi—Ying,HUANGXiao—Yuan,华中科技大学同济医学院附属同济医院肿瘤中心.湖北武汉430030CancerCenter,Ton~iHosp
2、ital,To,~iMedicalCoge.HuazhongUniversityofScienceandTechnology,Wuhan,Hubei,430030,P-R.China通讯作者:于世英Correspondenceto:YUShi—YingTel:86—27—83663342E—mail:syyu@tjh.tjmu.edu.cn收稿日期:2006—07.28修回日期:2006.12.11?基础研究?华.冷彦RNAlnterferenceonLineHeLaGAOQing—Lei,XIONGHua,LENGYan[ABSTRACT]BACKGROUND
3、&OBJECTIVE:Ku80isakeyproteinplaysarolejnrepairingDNAdoublestrandbreak(DSB)afterirradiation.ThereareafewstudiesaboutotherrolesofKu80exceptDSBrepair.ThisstudywastojnhibitKu80expressionjncervicaIcarcinomacellIineHeLawithsmallinterferingRNA(siRNA),andexploreitseffectoncellproliferati
4、on.METHODS:PlasmidspKu80一siRNAandpNeg—siRNA(negativecontro1)wereconstructedandtransfectedjntoHeLacells.TheexpressionofKu80jnHeLacellswasdetectedbyWesternblot.TheproliferationofHeLacellsinvitroand/nvivowasdeterminedbycloneformationassay,M1_rassay,andsubcutaneOustumorfOrmationjnnudemic
5、e.RESULTS:T^,ocelIcloneswerescreenedfrOmpKu80一siRNA—andpNeg—sIRNA—transfectedHeLacells.Ku80expressioninHeLacellswassuppressedmarkedlyaftertransfectionofpKu80一siRNA;thisclonewasnamedHela/Ku80.siRNA.ThecloneformationefficiencywassignificantlylowerinHeLa/Ku80一siRNAcellsthanincontrolcell
6、s(0.46+0.05vs.0.62+0.02,t=5.11,P<0.01).TheproliferationratewassignificantlyIowerinHeLa/Ku80?siRNAcellsthanjncontrolcellsat48hand72haftertransfection(P<0.05).Atthe25thdayaftersubcutaneoustransplantationjnnudemice,thetumorvolumewassignificantlysmallerjnHeLa/KuSO—siRNAgroupthaninc
7、ontrolgroup[(18.92±3.60)mmvs.(194.88~30.61)mm.,f=12.69,P<0.01].CONCLUSIONS:WesuccessfullyestablishedacellmodelthatKu80expressionjssuppressedalmostcompletelybysiRNAKuS0inhibitioninhibitstheproliferationofHeLacells『nvivoand/nvitro.KEYWORDS:Ku80;SmallinterferingRNA;Stabletransfection
8、:Cellprolife
