rail耐药的kasumi-1细胞株的建立及mrna表达谱差异分析

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1、数据处理找寻与TRAIL耐药可能相关的基因。结果：与Kasumi-1细胞相比，Kasumi-1R细胞表达上调大于2倍的基因有1537个，表达下调大于2倍的基因有487个。其中BCL-2家族的抑凋亡基因BCL2上调3.153倍，BCL2A1上调18.23倍，参与JAK/STAT通路的IFNAR1上调12.841倍，TRAIL的死亡受体TNFRSHOA表达下调3.256倍。结论：Kasum-1RTRAIL耐药可能与BCL2、BCL2A1和IFNAR1的表达上调、DR4表达下调有关。关键词：Kasumi-1细胞、TRAIL、耐药、DR4、IFNAR、BCL2、BCL2A1

2、2ABSTRACTBackground:Tumornecrosisfactorrelatedapoptosisinducingligand(TRAIL)iscytotoxictohumanleukemiacellswhileitisnotharmfultonormalcells.Butitsresistanceisalsouniversal.Researchonapoptosisoft(8;21)positiveacutemyeloidleukemiacelllineKasumi-1inducedbyTRAILshowedthesurvivalratenolonge

3、rdecreasedsignificantlywhenrsTRAILreachedacertainconcentrationwhichsuggestedKasumi-1cellswereresistanttoTRAILtosomeextent.PartITheestablishmentofTRAIL-resistantKasumi-1celllineObjective:ToestablishaTRAIL-resistantKasumi-1cellline(Kasumi-1R)andobservemorphology,andmolecularbiology;todet

4、ectmRNAexpressionofTRAILtranscriptvariants1-3andTRAILreceptors1-4intheparentalandresistantcellsandtheirexpressiononthetwocelllinessurface.Methods:1)Kasumi-1RcellswerederivedfromKasumi-1celllineintermittentlytreatedwithprogressivelyincreasedrsTRAILconcentration;2)Proliferationoftwocells

5、weremeasuredbyCCK-8assayandIC50oftwocelllinesandresistanceindexwerecalculatedaccordingtoproliferationoftwocellstreatedwithTRAILofdifferentconcentration.3)ThechangesofmorphologyoftwocelllineswereobservedundermicroscopeafterWright'sstaining.4)ApoptosiswasanalyzedthroughAnnexinV-FITC/PIdo

6、ublestainingbyflowcytometry.5)ThemRNAlevelofAML1-ETOfusiongene,ETOexon9a,TRAILtranscriptvariants1-3andTRAILreceptors1-4weredetectedbyRT-PCR.6)TRAILandTRAILreceptors1-4oncellsurfaceweredetectedbyflowcytometry.Results:1)Kasumi-1RcellsproliferationwasfasterthanthatofKasumi-1cells;2)24hour

7、sIC50forKasumi-1cellswas756.833ng/ml(logIC502.879±0.148)，24hoursIC50forKasumi-1Rcellswas1634646.005ng/ml(logIC506.213±0.637),theRIfor24hwas2159.48hoursIC50forKasumi-1cellswas345.390ng/ml(logIC502.538士0.153)，48hoursIC50forKasumi-1Rcellswas33642.641ng/ml(logIC50is4.257土0.317)，theRIfor4