RNA干擾EGFR基因?qū)β殉舶┘?xì)胞凋亡和細(xì)胞周期的影響.pdf

1、青島大學(xué)碩士學(xué)位論文RNA干擾EGFR基因?qū)β殉舶┘?xì)胞凋亡和細(xì)胞周期的影響姓名：張紅玲申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：婦產(chǎn)科學(xué)指導(dǎo)教師：陳愛平20090530TheEffectofRNAinterferencetoEGFRGeneoncellcycleandapoptosisofovariancancercelllineskov3AbstractobjectiveToconstructexpressionvectorofEGFRspecific

2、siRNAandtransfectitintohumanovariancancercelllineskov3ToinvestigatetheeffectofRNAinterferenceonthetargetgeneexpression，cellapoptosisandthecycleofthecelllineMethodsAccordingtoEGFRmRNAsequence，wedesignedandsynthesizedshRNA

3、targetedEGFRwhichweredirectlyclonedintopSilencerTM21一U6neoplasmidDNAsequencingconfirmedwhethertheinsertionsequencewascorrectornotThentransfectedtherecombinantplasmidintohumanovariancancercelllineskov3withlipofectamine200

4、0Therewerethreegroups：normalcontrol；non—specifictransfectandEGFRshRNAtransfectTheinhibitiveeffectofRNAionmRNAlevelandproteinlevelwererespectivelydetectedbyRTPCRandimmunocytochemistryAnnexinV／Pldoublestrainingbyflowc”omet

5、巧analysiswasusedtoevMuateapoptosisPIstrainingWaSconductedtoevaluatecellcyclearrestResultsPlasmidexpressivevectorsaimeddirectlyatEGFRweresuccessfullyestablishedEGFRsiRNAplasmidknockeddownEGFRexpressioninSkov3cellsobviousl

6、yFlowcytometryanalysisrevealedovariancarcinomacelllineSkov3sufferedanotableapoptosisandGdGtarrestafterRNAinterferencemediatedbysiRNAtargetingEGFRConclusionEGFRsilencingbyRNAicaninducecellapoptosisandarrestcellcycleatGo／G

7、iphase，whichshedslightonthepossibleuseofRNAiforfurtherinvestigationofthepathogenesisandgenetherapyofovariancarcinomaPostgraduatestudent：HongLingZhang(ObstetricsandGynecology)DirectedbyProfAiPingChenKeywords：ovarianneopla