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糖原合酶激酶3β以cyclin D1依赖性方式引发人肺腺癌细胞株A549细胞周期阻滞VIP免费

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204ActaPhysiologicaSinica,April25,2007,59(2):204-209http://www.actaps.com.cnReceived2006-11-23Accepted2006-12-29ThisworkwassupportedbytheNationalNaturalScienceFoundationofChina(No.30470757).*Correspondingauthor.Tel:+86-27-83692619;Fax:+86-27-83650729;E-mail:renliangwu@hotmail.comGlycogensynthasekinase3βinducescellcyclearrestinacyclinD1-dependentmannerinhumanlungadenocarcinomacelllineA549LIJian-Sha,ZHUMin,TIANDan,WANGMan-Xiang,WANGFang,LINa-Ping,WURen-Liang*DepartmentofPathology,TongjiMedicalCollege,HuazhongUniversityofScienceandTechnology,andKeyLaboratoryofPulmonaryDiseaseofMinistryofHealthofChina,Wuhan430030,ChinaAbstract:Theeffectofglycogensynthasekinase3β(GSK3β)hasbeenrepeatedlyimplicatedincellproliferation,butstudiesontheeffectofGSK3βindifferentcelllineswithdifferentstimulihavedrawndifferentconclusions.ToinvestigatethedirecteffectofGSK3βoncellgrowthinhumanlungadenocarcinomacelllineA549,wechangeditsactivitybytransienttransfectionwithtwokindsofGSK3βmutantplasmids,constitutivelyactiveformS9A-GSK3βanddominantnegativeformKM-GSK3β.Twenty-fourhourslater,cellcounting,flowcytometryandWesternblotdetectionweremaderespectively.TheresultsshowedthatenhancingGSK3βactivitycausedadecreaseincellnumber,aswellasahigherpercentageofcellsatG1phase.Further,theexpressionofcyclinD1wasdown-regulatedbyGSK3β.Takentogether,ourobservationssuggestthatGSK3βmayinduceG1cellcyclearrestinacyclinD1-dependentfashionandthereforepossiblyplaysagrowth-inhibitoryroleinA549cells.Keywords:glycogensynthasekinase3β;cellproliferation;cellcycle;cyclinD1糖原合酶激酶3β以细胞周期蛋白D1依赖性方式引发人肺腺癌细胞A549细胞周期阻滞李建莎,朱敏,田丹,王满香,王芳,李娜萍,吴人亮*华中科技大学同济医学院病理学系,卫生部呼吸系统疾病重点实验室,武汉430030摘要:对糖原合酶激酶3β(glycogensynthasekinase3β,GSK3β)在细胞增殖中的作用研究,在不同细胞系和不同刺激因素作用下得出了不同结论,本文旨在探讨GSK3β在人肺腺癌细胞系A549细胞生长中的直接作用。A549细胞瞬时转染持续激活型S9A-GSK3β以及显性负突变型KM-GSK3β两种GSK3β突变型质粒,改变GSK3β活性。24h后,分别进行细胞计数,流式细胞术及Westernblot检测。结果显示,增强GSK3β活性可导致细胞数量下降,G1期细胞百分比升高。细胞周期蛋白D1表达水平被GSK3β下调。结果提示,GSK3β可能以细胞周期蛋白D1依赖性方式引发A549细胞的G1期阻滞,从而发挥生长抑制效应。关键词:糖原合酶激酶3β;细胞增殖;细胞周期;细胞周期蛋白D1中图分类号:R322.3ResearchPaperGlycogensynthasekinase3β(GSK3β)isaserine/threo-nineproteinkinasethatwasfirstdescribedinametabolicpathwayforglycogensynthaseregulation[1].Incontrasttomanyotherkinases,GSK3βisconstitutivelyactiveinun-stimulated,restingcellsandbecomesinactivethroughphosphorylationatserine9byitsupstreamproteinkinases.Ithasavarietyofputativesubstrates,playingimportantrolesinmetabolism,cellproliferation,differentiationandsurvival[2].GSK3βattractsmoreandmoreattentionforitsrolesinadiverserangeofcellularprocessesanditskeypositionatseveralsignalingpathwaysthatarecrucialincancerandotherhumandiseases.Duringembryonichairfollicledevelopment,GSK3βmayserveasacentralsig-nalinghub,allowingfortightlyintegratedandspatiallycon-LIJian-Shaetal:GSK3βInducesG1CellCycleArrestinA549Cells205trolledproliferativeresponses[3].RecentdocumentsabouttumorspresentopposingeffectsofGSK3βoncellproliferation.Inastudyofcoloncancercells,inhibitionofGSK3βactivitybychemicalinhibitorsanditsexpressionbyRNAi...

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糖原合酶激酶3β以cyclin D1依赖性方式引发人肺腺癌细胞株A549细胞周期阻滞

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