雷帕霉素对hl-rapamycin to hl.docxVIP

EffectsandMoleculeMechanismsofRapamycinontheApoptosisofLeukemiaHL-60CellsAbstractobjective:ThisexperimenttakesacutepromyelocyticleukemiacellsHL-60asitsobjectandaimstoinvestigatetheeffectsofRapamycinontheapoptosisinductionofHL-60cellsandthemolecularmechanismsofapoptosis.Methods:TheinhibitationofHL-60cellsbyRapamycinwasmeasuredbyMTTassay.MorphicchangeoftheapoptosisofHL-60cellswasobservedbyinvertedmicroscopeandlightmicroscope,trapezoidstrapsbyDNAagarosegelelectrophoresis,andapoptosisratebyflowcytometry(FCM).Western-blotanalysiswasperformedtodetecttheexpressionofBaxandBcl-2proteinsafterrapamycintreatment.Results:RapamycininhibitedthegenerationofHL-60cellsdependingonitsconcentration.Theinhibitionofrapamycin80nmol/LequaledthatofATRA10nmol/L.HL-60cellsafterrapamycintreatmentof72hourswereobservedtomaketypicalapoptosistransformation,suchasdiminution,karyopycnosis,etc,throughlightmicroscopeandAO/EBFluophot.DetectionofagarosegelelectrophoresiswasthatDNAtrapezoidstrapswereseenindistinctlywhenthedensityofrapamycinwas40nmol/L,butclearlywhenthedensityincreasedto80nmol/L.However,whenthedensityrisedto160nmol/L,hymenodecelldeathstrapswerealsoseen.Inductionapoptosisofrapamycin80nmol/LonHL-60cellswasdetectedthroughflowcytometry,andwouldnotincreasewithrisingdensity.Itwasafter72hoursofRapamycin80nmol/Ltreatmentthatdistinctapoptosiswasobserved.Resultofwestern-blot:Withtheextensionoftime,Bax/Bcl-2wasup-regulatedwithriseofapoptosisalbumenBaxandfallofBcl-2.Conclution:RapamycininhibitedtheproliferationofleukemiaHL-60cellsdependentondensityandtime.Theapoptosismechanismmaybeassosiatedwiththeup-regulationoftheratiobetweenBaxandBcl-2.Postgraduat:YalanCuiDirectedbyPro.ShiyunCaoInstituteofPhysiology,NanhuaUniversityHengyang,421001KeyWords:Rapamycin,leukemia,proliferation,apoptosis前言根据WHO的报告，2000年全球癌症死亡人数已经超过700万，占全部死亡人数的12%，在发展中国家占9%，在发达国家占21%。2005年全球有760万人死于癌症。如果这一趋势得不到改善，预期到2020年每年新发生的癌症患者将达1500万，在发展中国家癌症总数将增加73%，发达国家为29%。我国是发展中国家，总体来说，癌症占居民死亡原因的19%，居常见死亡原因的首位，已经接近发达国家。白血病是一类源于造血干细胞的恶性疾病,是儿童中最常见的