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ID:31483739
大小:81.00 KB
页数:15页
时间:2019-01-11
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1、WORD完美整理版ExpressionofliviningastriccancerandinductionofapoptosisinSGC-7901cellsbyshRNA-mediatedsilencingoflivingeneBackground-Becauseofincreasedresistancetoapoptosisintumorcells,inhibitionofspecificantiapoptoticfactorsmayprovidearationalapproachforthedevelopmentofnoveltherapeuticstrategies.L
2、ivin,anovelinhibitorofapoptosisproteinfamily,hasbeenfoundtobeexpressedinvariousmalignanciesandissuggestedtohavepoorlyprognosticsignificance.However,nodataareavailableconcerningthesignificanceofliviningastriccancer.Inthisstudy,wedetectedtheexpressionoflivininhumangastriccarcinomaandinvestigat
3、edtheapoptoticsusceptibilityofSGC-7901cellbyshRNAmediatedsilencingofthelivingene.Methods-ThemRNAandproteinexpressionoflivinwereanalyzedbyRT-PCRandwesternblotassay.Therelationshipbetweenlivinexpressionandclinicalpathologicparameterswasinvestigated.ThesmallinterferingRNAeukaryoticexpressionvec
4、torspecifictolivinwasconstructedbygenerecombination,andthenucleicacidwassequenced.ThenitwastransfectedintoSGC-7901cellsbyLipofectamin2000.RT-PCRandWesternblotassaywereusedtovalidategene-silencingefficiencyoflivininSGC-7901cells.StablecloneswereobtainedbyG418screening.Thecellapoptosiswasasses
5、sedbyflowcytometry(FCM).Cellgrowthstateand50%inhibitionconcentration(IC50)of5-FUandcisplatinwasdeterminedbyMTTmethod.Results-TheexpressionoflivinmRNAandproteinweredetectedin19of40gastriccarcinomacases(47.5%)andSGC-7901cells.Noexpressionoflivinwasdetectedintumoradjacenttissuesandbenigngastric
6、lesion.Thepositivecorrelationwasfoundbetweenlivinexpressionandpoordifferentiationoftumorsaswellaslymphnodemetastases(P<0.05).FoursmallinterferingRNAeukaryoticexpressionvectorspecifictolivinwereconstructedbygenerecombination.Andoneofthemcanefficientlydecreasetheexpressionoflivin,theinhibition
7、ofthegenewasnotlessthan70%(P<0.01).Therecombinatedplasmidswereextractedandtransfectedgastriccancercells.ThestablecloneswereobtainedbyG418screening,andwereamplifiedandcultured.Whenlivingenewassilenced,thereproductiveactivityofthegastriccancercellswa
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