FMDV 2A多順反子溶瘤腺病毒在肝癌治療中的研究.pdf

1、Y10二2235尚≯理乒大學ZhejiangSciTechUniversity碩士學位論文論文題目E蛆瞼z2叢壘19垂縋齷盤邀邋耋舡盤造矗立曲紐疊——q舉t。專業(yè)：生壁塑型重量整皇塑星一一作者姓名：指導教師：完成日期：一劉監(jiān)i蘭一盛讎魚!緝型L浙江理工大學碩士學位論文ABSTRACTCancersofteninvolvedisordersinmultiplecellulargenes，thetreatmentofwhichmaynece

2、ssitatethedeliveryofmultipletherapeuticgenesintothecells，Anditiswellacceptedthatacombinationapproachsimultaneouslytargetingmultipledefectivegeneswouldbeexpectedtobemoreeffectivethansingle—genestrategySeveralstrategieshav

3、ebeendescribedtoco—expresstwoormoregenes，includingsplicing，fusionproteins，reinitation，internalpromoters，multiplepromotors，monocstronic，internalribosomeentrysite(IRES)，selfcleavingpeptide2A，etc哆Nevertheless，mostoftheseapp

4、roacheshavesignificantdrawbacks，whichgreatlylimittheirapplicationsingenetherapysincetheexpressionleveloftransgenesoftendramaticallyaffectsthetherapeuticefficacyThelargesizeandtheimbalanceofmostIRESeswhichmakeitverydiffic

5、ulttopredicttheexpressionlevelofthedownstreamcistron，andgreatlyblocktheirwideemploymentinmultiplegenestherapystrategyAnalternativestrategytoguaranteeareliableCO—expressionwasthereforeintroducedFMDV2Ahasbecomeanattractive

6、elementingenetherapyforcoordinateco—expressiontwoormultipletherapeuticgenesThesingle，long，openreadingframe，IL一24—2A—TRAIL，wasincorporatedintotheE3regionofadualtargetingoncolyticadenovirusAdCN204，anadenovimsengineerdtotar

7、getbothofTERTandRbpathways，andundcrthecontrollingoftheE3endogenouspromotorSeveralcelllineswereinfectedwiththeadenovimstoinvestigatethe‘cleavage’activityof2AWesternBlottingwereperformedtodetectthetranslationproductsThreek

8、indsoftranslationproducts，IL一24—2ATRAIL，IL一24—2AandTRAIL，weredetectedincellstreatedwiththisbicistronicadenovirnsthroughWesternBlotting，andfurtherquantifiedoninfraredimagingsystemAbout50％～75％oftranslationproductswereinthe

9、cleavedformandwealsofoundallexcessofthetranslationproductofNterminalof2A(IL一24—2A)withrespecttotheC—terminalof2A(TRAIL)，andtheII。24—2A：TRAILvariedfrom6：1to2：1indifferenttissues，andfrom3：1to2：1inlivertissueOurstudyshowedf

10、orthefirsttimethatFMDV2AhadhighcleavageactivityinouradenovirnsandwasapromisingstrategytoachievepotycistronicsystemsTheanitumorefficacyofthenovelreplicativeadenoviruswasfurtherinvestigatedinexperimentalHCCTreamaentwithAdC