苯甲醇对红细胞冻干前海藻糖负载红细胞影响研究 (2).doc

阿霉素对K562细胞Gfi1表达的影响及其与相关凋亡基因关系的研究
作者：常伟孙汉英黄敏周剑锋张义成作者单位：华中科技大学同济医学院附属同济医院血液科，武汉430030
【摘要】本研究探讨阿霉素体外作用于K562细胞的细胞效应及癌基因Gfi1和相关凋亡基因表达变化的机制。用不同浓度的阿霉素作用于K562细胞24小时,然后应用DNA电泳和流式细胞术检测K562细胞凋亡;用RTPCR、流式细胞术检测Gfi1、Bcl2、bax基因和蛋白表达的变化。结果表明：当阿霉素浓度在0、0.5、2.0mg/L作用K562细胞24小时,细胞凋亡增加,可见典型的DNA断裂电泳条带;同时,当ADM浓度自0.5增为2.0mg/L时,Gfi1表达减少,bax表达增加;Bcl2表达在ADM0.5-2.0mg/L之间变化不明显,mRNA及蛋白水平均无统计学差异；当阿霉素浓度大于2.0mg/L时,细胞凋亡率并不增加,而是下降,且出现细胞坏死。结论：一定浓度阿霉素能诱导K562细胞凋亡,细胞凋亡率表达的变化与阿霉素浓度呈一定的量效依赖关系,阿霉素诱导K562细胞凋亡可能与抑制Gfi1基因的表达和激活bax基因的表达有一定的关系。
【关键词】阿霉素K562细胞；细胞凋亡；Gfi1；Bcl2；bax
EffectofAdriamycinonGfi1ExpressioninK562CellsandItsRelationshipwiththeRelevantApoptoticGenesCHANGWei,SUNHanYing,HUANGMin,ZHOUJianFeng,ZHANGYiChengDepartmentofHematology,TongjiHospital,TongjiMedicalCollege,HuazhongUniversityofScienceandTechnology,Wuhan430030,China
AbstractThestudywaspurposedtoexploretheeffectofadriamycin(ADM)onK562cellsinvitroandthemechanismofexpressionchangesofrelevantapoptoticgenesandoncogeneGfi1.Theapoptosiswasassayedbyflowcytometry(FCM)andtheDNAelectrophoresis;theexpressionchangesofGfi1、Bcl2、baxmRNAandproteinweredetectedbyRTPCRandFCMafterK562cellsweretreatedwithdifferentconcentrationsofADMfor24hours.TheresultsshowedthatwhenK562cellsweretreatedwith0-2.0mg/LADMfor24hours,thetypicalapoptoticDNAelectrophoresisbandofK562cellswereobservedwiththedoseincreasing.WhenconcentrationofADMwas0.5and2.0mg/L,theexpressionofGfi1decreasedandtheexpressionofbaxincreased;whenconcentrationofADMwas0.5-2.0mg/L,theexpressionofBcl2wasnotfoundtobesignificantlychanged,thelevelsofBcl2mRNAandproteinwereofnostatisticaldifference.WhendoseofADMwashigherthan2.0mg/L,thepercentageofapoptoticK562cellsdecreasedwithcellnecrosis.Itisconcludedthatatcertainrangeofconcentration,apoptosisornecrosisofK562cellscanbeinducedbyADM,thepercentageofapoptosis,thechangesofexpressionofBcl2,baxandGfi1dependonthedoseofADM.ThemechanismofapoptosisinK562cellsinducedbyADMmayberelatedtosuppressionofGfi1oncogeneandactivationofexpressionofbaxgene.
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