苯甲醇对红细胞冻干前海藻糖负载红细胞影响研究.doc

SODD和Survivin在化疗药物诱导白血病细胞凋亡中的作用
作者：陶红芳,胡群,方建林,刘爱国,刘双又,张柳清,胡迎作者单位：华中科技大学同济医学院附属同济医院儿科,武汉430030；1华中科技大学同济医学院附属协和医院放射科，武汉430022
【摘要】为了研究死亡结构域沉默子（SODD）、survivin、caspase3、caspase8及caspase9在化疗药物诱导白血病细胞凋亡中的变化，以进一步明确凋亡调控基因的调控机制和寻找药物作用新靶点，采用AnnexinV/PI双标记流式细胞术检测化疗药物作用后Jurkat细胞凋亡发生率；采用免疫印迹法分析SODD、caspase3、caspase8及caspase9蛋白表达的变化；采用RTPCR检测survivinmRNA表达的变化；采用免疫组织化学SABC法检测survivin蛋白表达的变化。结果表明：SODD及survivn高表达均抑制肿瘤细胞凋亡，VCR具有时间依赖性特异下调SODD蛋白的功能并能有效诱导Jurkat细胞凋亡，在该凋亡过程中caspase3、caspase8酶原呈时间依赖性逐渐被水解剪切，而caspase9及survivin均无明显变化趋势。结论：SODD参与了VCR诱导白血病细胞凋亡过程,VCR通过下调SODD表达并启动受体配体介导途径诱导Jurkat细胞凋亡，在该过程中线粒体凋亡途径并未被启动。
【关键词】白血病SODD；survivin化疗药物细胞凋亡
EffectsofSODDandSurvivinonLeukemiaCellApoptosisInducedbyChemotherapeuticDrugsTAOHongFang,HUQun,FANGJianLin1,LIUAiGuo,LIUShuangYou,ZHANGLiuQing,HUYingDepartmentofPediatrics,TongjiHospital,TongjiMedicalCollege,HuazhongUniversityofScienceandTechnology,Wuhan430030,China;1DepartmentofRadiology,UnionHospital,TongjiMedicalCollege,HuazhongUniversityofScienceandTechnology,Wuhan430022,China
AbstractThisstudywasaimedtoinvestigatethechangesofsilencerofdeathdomains(SODD),survivin,caspase3,caspase8andcaspase9intheapoptoticprocessofhumanleukemiacellsinducedbychemotherapeuticdrugsinordertoexplorethemolecularmechanismofapoptoticmodulatorygenesandtosearchforthenewtargetofchemotherapeuticdrugs.AfterJurkatcellswereinducedbychemotherapeuticdrugs,thetranslocatedphosphatidylserinewaslabeledwithannexinV/PI,andtheapoptosisincidencewasmeasuredbyFCM;TheexpressionchangesofSODD,caspase3,caspase8andcaspase9weredeterminedbyWesternblot;thechangesofsurvivinmRNAandproteinweredeterminedbyRTPCRandimmunohistochemistrySABCmethodrespectively.TheresultsindicatedthathighexpressionsofSODDandsurvivincouldinhibitapoptoticsignalingpathway;VCRdownregulatedthefunctionofSODDproteinandeffectivelyinducedtheapoptosisofJurkatcellsinatimedependentmannerandactivatescaspase3throughthedeathreceptormediatedactivationofcaspase8,inwhichcaspase9andsurvivinwerenotdegraded.ItisconcludedthatSODDp