GCV靶向抗肝癌及旁观者效应的研究的开题报告.docxVIP

AF-hTERT-TK/GCV靶向抗肝癌及旁观者效应的研究的开题报告

（Title）：StudyontheTargetedAntitumorEffectandbystandereffectofAF-hTERT-TK/GCVinLiverCancer

（Introduction）：

Livercancerisoneofthemostcommonmalignanttumorsintheworld.Althoughtraditionaltreatmentssuchassurgery,chemotherapy,andradiotherapyhaveachievedcertaineffects,therearestillmanylimitations,suchasdrugresistance,tumorrecurrence,andseveresideeffects.Therefore,itisnecessarytofindnewandeffectivetreatmentmethods.

AF-hTERT-TK/GCVisatargetedgenetherapythathasshownpromisingresultsinpreclinicalstudies.TheAF-hTERT-TKgenecanspecificallytargettelomerase-positivelivercancercells,whileTKconvertsGCVintoatoxicmetabolite,causingcelldeath.Furthermore,thebystandereffectcanenhancethetherapeuticeffectbycausingneighboringcellstoundergoapoptosis.

（Objectives）：

TheaimofthisstudyistoinvestigatethetargetedantitumoreffectandbystandereffectofAF-hTERT-TK/GCVinlivercancer,includingthefollowingobjectives:

1.ConstructtheAF-hTERT-TKexpressionvectorandperformtransfectioninlivercancercellstoverifyitsspecificityandefficacy.

2.ObservetheapoptosisandcellcyclechangesinAF-hTERT-TK/GCV-treatedlivercancercellsandexplorethepossiblemechanism.

3.EvaluatetheinvitroandinvivoantitumorefficacyofAF-hTERT-TK/GCV.

4.InvestigatethebystandereffectofAF-hTERT-TK/GCV,includingitsextentandtheunderlyingmechanism.

（Methods）：

1.ConstructionoftheAF-hTERT-TKexpressionvectorandtransfectionintolivercancercells.

2.DetectionofthegeneexpressionusingRT-PCRandWesternblotting.

3.ObservationofapoptosisandcellcyclechangesinAF-hTERT-TK/GCV-treatedlivercancercellsusingflowcytometry,TUNELstaining,andHoechst33342staining.

4.EvaluationoftheantitumorefficacyofAF-hTERT-TK/GCVinlivercancercellsandaxenograftmodelusingMTTassay,colonyformationassay,andtumorg