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ID:28573609
大小:9.24 MB
页数:51页
时间:2018-12-11
《rhoa蛋白在人胃癌细胞株sgc7901的定位和移位》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、江苏大学硕士学位论文结果:(1)内源性的RhoA在人胃癌细胞株SGC.7901的细胞膜、细胞浆和细胞核中均有分布。(2)外源性的RhoA在人胃癌细胞株SGC.7901的细胞膜、细胞浆和细胞核中均有分布。(3)在人胃癌细胞株SGC一7901的间期细胞核中,RhoA定位在核仁区。(4)在人胃癌细胞株SGC.7901中,RhoA基因没有发生碱基对突变。(5)LPA能够引起RhoA从细胞浆向细胞膜和细胞核移位;CPT-cAMP能够引起RhoA从细胞膜和细胞核向细胞浆移位;CPT-cAMP能够阻断LPA引起的RhoA移位。结论RhoA基因在人胃癌细胞株SGC.7901中没有
2、发生突变。RhoA蛋白在SGC.7901细胞的细胞膜、细胞浆和细胞核中均有分布,在间期细胞核1为RhoA定位于核仁区;LPA能够活化RhoA,使其向细胞膜和细胞核移位,CPT-cAMP能够抑铝IJRhoA的活性,使其向细胞浆移位,CPT-cAMP能够阻断LPA弓[起的RhoA在细胞内的移位。关键词:RhoA,定位,移位,核仁,LPA,CPT-cAMPⅡ江苏大学硕士学位论文ABSTRACTobjectiveToinvestigatethesubceUularlocalizationofRhoAinhumanGastricCancerCellLineSGC-7901a
3、ndtoelucidatetheeffectsofactivitychangeofRhoAonitstranslocationwithinthecells.Methods(1)Immunofluorescenceassay,immunocytochemistryassayandimmuno—TEMassaywereappliedfordetectingthelocalizationofendogenousRhoAinhumanGastricCancerCellLineSGC-7901.(2)WesternBlotassaywasappliedfordetectin
4、gendogenousRhoAindifferentsubcellularfractions(membrane,cytosolandnucleusfractions).(3)HumanGastricCancerCellLineSGC一7901cellsweretransfectedwithplasmidDNAencodingdifferentstructuralRhoA(wt,63LandN19),andWesternBlotassaywasappliedfordetectingexogenousRhoAindifferentsubcellularfraction
5、s(membrane,cytosolandnucleusfractions).(4)RhoA(wt,63LandN19)geneswereconstructedintopcDNA3vectorscontainingGFPgeneandtheplasmidsweretransfectedintohumanGastricCancerCellLineSGC一7901cells.ThelocalizationofexogenousRhoAinlivingcellswasinvestigatedbylaserscanningconfocalmicroscope.(5)Hum
6、anGastricCancerCellLiIleSGC-7901cellsweretreatedwithHoechst33342forrevealingchromatin,andimmunofluorescenceassaywasappliedforinvestigatingthepreciselocalizationofRhoAinnucleus.(6)WesternBlotassaywasappliedfordetectingRhoAinnucleolifractionfromhumanGastricCancerCelllineSGC.7901cells.(7
7、)ThefulllengthofRhoAgenewasclonedfromhumanGastricCancerCelllineSGC一7901cellsandthesequencewasanalyzed.(8)WesternBlotassaywasappliedforinvestigatingthetranslocationofⅢ江苏大学硕士学位论文RhoAinhumanGastricCancerCelllineSGC一7901cellsinducedbyLPAandCPT—a~MPrespectively,andtheeffectofCPT-a刘MPonLI'A
8、—indu
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