靶向乳腺癌的增殖腺病毒載體的研究.pdf

1、南京師范大學碩士學位論文靶向乳腺癌的增殖腺病毒載體的研究姓名：胡小翠申請學位級別：碩士專業(yè)：細胞生物學指導教師：蘇長青曹祥榮2009南京師范大學碩士學位論文IIABSTRACTOncolyticadenovirusesarecurrentlybeingdevelopedasanticancerbiologicalagents.Creatinganarmedtherapeuticviralplatfmwherethelyticproper

2、tyofviralinfectioncoupledtoviralmediatedtransgenedeliverymayenhanceviralbasedtherapeuticpotential.Asoncolyticadenovirusspecificallyproliferatingreplicatingintumcellstheanticancergenesmayhaveasubstantialincreaseincopyitse

3、xpressionlevelmayrisehundredsoftimesconsequentlyachievethepurposeofkillthetumcells.InthisresearchamodifiedbreastcancerspecificpromotermHerDF3wasclonedusedtodrivetheexpressionofmutatedE1A(whichwasdtheCR2regionlosttheabili

4、tyofcombiningwithRb)mutatedE1B(whoseE1B55KDwasdefectivelosttheabilityofbindingtop53).HereatumspecificreplicativevectpSCQwasconstructedinwhichthemutatedE1AE1BwascontrolledbymHerDF3promoterthenpSCQhIL24wasconstructedbyingt

5、hehIL24geneintopSCQ.TherecombinantadenoviruseswerepackedinHEK293namedAdSCQAdSCQhIL24respectively.TheadenoviruseswereidentifiedbyPCRtheninfecteddifferentcelllines.TheivereplicationofAdSCQAdSCQhIL24intumcellswasinvestigate

6、dbyvirusproliferativetestWesternblotassayCPEassaywhileinnmalcellswasnotobvious.TheseresultsindicatedthatanovelcancerivegenetherapeuticoncolyticadenovirusvectsystemnamedAdSCQwasconstructedwhichcanivelyreplicateexpressther

7、apeuticgenesincancercells.Itisapromisingsystemftargetedcancergenetherapy.InadditionweconstructedarecombinantprokaryoticexpressionvectofhIL24expleditsproperinducingconditions.Allthewksarehelpfulfitsexpressionpurificationf