rhoa蛋白在人胃癌细胞株sgc7901的定位和移位

《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