重组分泌型strail腺病毒的构建与其诱导脑胶质瘤u251细胞凋亡的实验的研究

《重组分泌型strail腺病毒的构建与其诱导脑胶质瘤u251细胞凋亡的实验的研究》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、AbstractobjectiveT。c。nstnlctasecretingrec。mbinantadenovirusVectorwiththeeXtracenularregionoftheh啪anTRAILgene(sTRAIL)andtoinvestigatetheantitumoractiVities。fAd-sTRAIL0nmalignantgliomacelllineU251·MethodsTheTRAILgene脚entencoding14-281aminoacidsfromhumanlivercDNAI．b删was跚plifiedbyPCR．mpAdTrack

2、-CMV-IL．2-TRAILwasconstructedbYins叭ingTRAILintotheadenoviralshuttlevectorpAdTraek．CMV-IL-2withsecretingpeptidesIL-2．PmeI-digestedpAdTrackCMV-IL．2．TRAILwaStransformedintoE．coliBJ5183harboringpAdEasy-1byelec仃0transfomation．RecombinantplasmidpAd-sTKAILwasselected·PackingintheHEK293cells，there

3、l，lication-deficientadenovinlsAd-sTRAlLparticleswaspurifiedbycesiumchloridedensitygradientcentrifugationanddetectedtheti溉ThroughRT-PCRandwetembIo仳ing，meeXpre5slonot伽翟etgenewaSdetected．CelllineU251wasinfectedbyAd—sTRAIL·TheexpressionofsTRAILw舔detectedbyimmunofluorescence．Cellviabilitywasdet

4、erminedbyMTT·Turnorcell8印opt0818嗍demonstratedbymot'l：Ihologicchanges，flowcytometry．Cellsofsub‘phasesandcellcyclewe陀dete酏edbyflowcytometrywithPIstaining’R髑uItsThegenefrag_ment0f104bpwasamplifiedbyRT-PCR·Theexpressionofl鹕etgenemu25lcellsinfectedbyAd．sTRAILwasshownbywestemblottingandimmunoflu

5、orescence·mhighe。rate0fap。咖siswaSdemonstratedbyMTTmethod，flowcytometry．Therate。fAd。sTRAlLinducedu251ce¨apoptosismasedwiththedosageandtimeincrease．TheapoptosisofG0／Gl，sp№celIswe心mo舱significantthanthecontrolgroups·c。眦IusionThesecretingrecombinantaden。viralvectorwiththeextraeellularregion0fhu

6、manTRAILgeneh邪beenconstructcdsuccessfully．Thegrowthandproliferati。nofU251cell1inew舔inhmitedmertheinfectionofAd．sTRAIL．Itisconcludedthatap。ptosiscoiledthecellcycleprog删on锄dprcsentacellcyclespecificity，implyingthatapoptosisinducedbysTRAILisacellcycleeVent·Postgraduate：Yah"gangWangDirectedby：

7、Prof．YiheDouKeyw。rd：TRAIL；recombinantadenovi吣gli。ma；apopt。sis；ceu锣cle引言2010年ESMO(EuropeanSocietyforMedicalOncology)关于高级别胶质瘤的临床诊疗指南指出，目前脑胶质瘤的发病率达～5／100000，发病人群覆盖所有年龄段，其中以15周岁到60周岁之间发病率最高Ⅲ。胶质瘤是最常见和死亡率最高的神经系统肿瘤，大约占原发性中枢神经系统肿瘤的80％。按照WHO分级，脑恶性胶质瘤包括：胶质母细胞瘤(WHOⅣ级)，间变