急性髓系白血病的分子诊断分析-molecular diagnosis analysis of acute myeloid leukemia.docx

急性髓系白血病患者IKAROS6表达研究目的：本研究旨在探讨急性髓系白血病患者IKAROS6表达情况及临床特征。方法：采用PCR扩增产物克隆测序法检测205例急性髓系白血病(AML)患者中IKAROS6表达情况；结合患者临床及实验室资料进行统计学分析，探讨IKAROS6表达与患者临床资料及预后之间的相关性；表达谱芯片聚类分析IKAROS6表达的AML-M5患者与无IKAROS6表达AML-M5患者组之间的差异基因。正常对照为10名与患者无血缘关系的健康志愿者。所有的患者和志愿者均经知情同意。结果：10例健康对照标本中未检测到IKAROS6表达。205例AML中23例(11.22%)出现IKAROS6表达，其中M4/M5表达IKAROS6阳性率为(17/86,19.77%),M1表达IKAROS6阳性率为(1/18,5.56%)，M2表达IKAROS6阳性率为(3/71,4.23%)，M3表达IKAROS6阳性率为(2/28,7.14%)。在有完整统计数据的109例AML患者分析显示IKAROS6表达阳性的患者完全缓解CR率(6/23,26.09%),显著低于IKAROS6表达阴性组(65/92,70.65%)(P＜0.01)；IKAROS6表达与性别、NPM1、CEBPα、C-kit、IDH1、IDH2、FLT3突变无明显相关性(P＞0.05)；与DNMT3A突变以及MLL表达有极显著相关性(P＜0.01)。采用GEP聚类分析将IKAROS6阳性组、IKAROS6阴性组及MLL阳性组比较，分析图显示，IKAROS6阳性组有独特的基因表达模式。结论：IKZF1基因异常表达可见于AML，以IKAROS6异构体为主，多见于AML-M4，AML-M5；在AML患者中IKZF1异常表达是独立的预后不良指标；聚类分析提示IKAROS6阳性AML具有独特的表达谱，或可能成为AML的又一个亚型，为AML诊断和临床治疗提供新的靶点。【关键词】IKZF1；IKAROS；急性髓系白血病；逆转录聚合酶链反应AbstractTheroutinegeneticsexaminationofacutemyeloidleukemiaPurpose:Toscreenthegeneticaberrationsofdenovoacutemyeloidleukemiausingcytogeneticandmolecularcytogenetictechniques.Methods:Mononuclearcellsacquiredfromthebonemarrowof205patientswithdenovoacutemyeloidleukemiawerecollected,thebasicclinicaldatawererecordedaccordingly.KaryotypewasanalyzedusingG-bandingofchromosomes.FusiongenesandinternaltandemduplicationofFMSliketyrosinekinase3(FLT3-ITD)wasdetectedbypolymerasechainreaction.Mutationsofnucleophosmin(NPM1),CCAATenhancerbindingfactorα(CEBPα)andC-kitgenesweredetectedbydirectsequencingofPCRproduct.Theclinicaldatawereanalyzedwiththeresultsofgeneticscreening.Results:Kayrotypewasanalyzedin166cases;resultsoffusiongeneswereavailablein202cases.FLT3-ITDmutationwasfoundin45patients;NPM1mutationwasfoundin34patients;CEBPαmutationwasfoundin61patients;C-kitmutationwasfoundin16patients.Theformerthreewereallassociatedwithnormalkaryotype,butwithdifferentFABsubtyperespectively.TheC-kitmutationwasassociatedwitht(8;21)translocation.Conclusion:Thecombinationofcytogeneticandmolecularcytogenetictechniquescanhelptotellthebackgroundofpatientswithdenov