survivin, caspase-3在皮肤鳞状细胞癌中的表达及雄黄的体外干预分析

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1、ExPressionofSurvivin，CasPase一3ProteinsincutaneoussquamouscellcarcinomaandinterventioninvitrobyRealgarsPeeialty:CliniealeombinationofTCMwitllwesternmedieineAuthor:Zhangehun一minSuPervisor:ProfessorQi州an一允Abstractobjeetives:Survivin，amemberoftheinhibitorofaPoPtosisProtein(IAP)几mil又eandireetlyinL

2、hibitCasPase一3andCasPase一7aetivityandPlayanimPortantroleinoneogenesis.ThisstudywasdesignedtoinvestigatetheexPressionofSurvivinandCasPase一3incutaneoussquamousee]1careinoma(SCC)andinvestigatethesignifieaneeandrelationshipofSurvivinandCasPase一3inthepathogenesisanddevelopmentofsquamouseelleareino

3、ma:InthisstudywetrytoexaminewhetherRealgarhasaneffeetonhumanCutaneousSquamousCellCarcinomaCellLineA431ininhibitingPro]iferationandindueingaPoPtosisbyMT工FCMandRl’-PCRinordertoProvidetheoryandexPerimentbaSC.Methods:(l)TheexPressionofSurvivinandCasPase一3wasdeteetedtouseimm朋ohistochemiea]staining

4、in46SPeeimensfromSCCPatients;20normalskintissueswereusedaseontrol.(2)Cellculture:A43]cellswereeulturedinDMEMmediumeontaining10%fetalealfserum(FCS)，Penieillin(10Ou/ml)，andstrePtomyein(IO0u/ml)，in5%eoZ，at37，Csaturationhumsditybox.whentheeellsgrewtoformamonolayer，theywerestimulatedWithRealgarfor

5、24hoursandrhenbeeolleeted.(3)MTT:A431eellswereineubatedinthe96一welleultureboard.Culturfluideontainswith30林g/ml，60林g/ml，100林g/ml，200拼g/ml，400林g/m]Realgar，whieheontrasttotheeulturemediumcellsWithoutRealgar.EaehwelladdMTTsolution(5“g/ml)reagent20ml，toeulture4h，at37oC，andstopeulture.EaehwelladdDM

6、SO150林l，eonfusing10min，usingELISAreadertodeteettheA49Oatwavelength490nmofeaehwell.Restrainrate(%)=!l一A49o(thedisPosalteam)/A49。(theeontrastteam)xl00%].(4)Cellmorphologyobservation:WiththeeulturefluidinwhiehtheeoneentrationofRealgarwas30一200林g/ml，observingtheeellmorphologyandtakingPhotoswithin

7、vertedPhaseeontrastmieroseroPeafter24hculture.(5)FCM:xneubatingA431cellsin6一welleulturemediumwith1x106/wel一，a且er24heuzturewiththeeoneentrationofRealgar30一200拼g/ml，gatheringeellsgrowingalongthewal]andmakingmonocellularSuspension，andthenadds林1A