Apoptosis of bladder transitional cell carcinoma T24 cells induced by adenovirus-mediated inducible nitric oxide synthase gene transfection

(整期优先)网络出版时间:2013-05-15
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Objectives:Toinvestigatetheeffectsofadenovirus-mediatedinduciblenitricoxidesynthasegenetransfectiononbladdertransitionalcellcarcinomaT24cells,andtoprovidenovelinsightsandapproachestoclinicaltherapiesagainstbladdertransitionalcellcarcinoma.Methods:Firstly,constructrecombinantadenovirusvectorpAd-iNOSofiNOS,followedbytransfectionofpAd-iNOSintoHECK293packagingcells.Thirdly,harvestrecombinantadenovirusrAd-iNOSafteramplificationandpurificationprocedures.Finally,transfecttherecombinantadenovirusrAd-iNOSintohumanbladdercarcinomaT24cellsandexaminetheeffectofrAd-iNOStransfectiononapoptosisofT24andpossiblemechanism.Results:Asshownbythisstudy,therecombinantadenovirusrAd-iNOSwasconstructedsuccessfully.Thevirustiterwas5.8×108PFU/mLandrecombinantwasverifiedbyPCRanalysis.TransfectionofadenovirusrAd-iNOSintoT24cellscouldinducesecretionofhighNOconcentration,P53proteinexpressionupregulation,aswellaspromotionofT24cellapoptosis.Conclusions:ThetransfectionofhumanbladdercarcinomaT24cellsfromrecombinantadenovirusrAdiNOSwasconfirmedtoinduceintracellulariNOSover-expression,highproductionofNO,up-regulationofintracellularP53expressionandpromotionofcellapoptosis.