去泛素化酶UCH—L3和自噬基因LC3 Ⅱ在急性髓细胞白血病诊断中的临床应用研究  

The role of UCH-L3 and LC3 II in the diagnosis of acute myeloid leukemia

作　　者：王楠[1] 胡丹 袁宏[1] 

机构地区：[1]大连医科大学附属第一医院检验科,116023 [2]大连医科大学

出　　处：《中华检验医学杂志》2018年第1期53-58,共6页Chinese Journal of Laboratory Medicine

基　　金：辽宁省科学技术计划项目（2015020322）;大连市科技局项目（2015E12SF166）

摘　　要：目的探讨自噬和UCH－L3在急性髓细胞白血病发生发展过程中的作用机制和相互关系，为白血病的诊断及治疗提供新的靶点和策略。方法选取2014至2015年大连医科大学附属第一医院收治的84例AML患者（男47例，女37例）及健康体检对照组25名（男13名，女12名）。AML初发组40例（男24例，女16例），中位年龄54岁（23～85岁）；缓解组30例（男14例，女16例），中位年龄45岁（22～74岁）；难治组14例（男9例，女5例），中位年龄50岁（18～80岁），其中M1型3例、M2型42例、M3型18例、M4型3例及M5型18例。HL－60细胞采用Real Time PCR及Western Blot法检测TCID处理前后UCH－L3和LC3II的表达变化。AML患者及对照组采用Real Time PCR方法检测外周血单个核细胞UCH－L3和LC3II基因mRNA的表达情况；比较其在初发组、缓解组、难治组及对照组间表达情况；分析AML不同型别之间UCH－L3和LC3II的表达水平；比较AML初发组LC3Ⅱ基因表达的高低与临床特征、临床分期、实验室检查结果及治疗效果的关系。追踪同一患者治疗前后UCH－L3及LC3ⅡmRNA的表达变化。计量资料用t检验，率的比较用χ2检验；相关性分析采用秩相关检验；不符合正态分布的数据用非参数检验。结果TCID处理HL－60细胞后，UCH－L3及LC3 II在基因和蛋白水平上与未处理组相比表达均下调，差异有统计学意义（t＝－29.435及－8.105，P均〈0.05）；AML初发组的UCH－L3表达量较健康对照组下调，差异有统计学意义（Z＝－3.87，P〈0.05）；AML缓解组UCH－L3表达量与初发组和难治组相比均高表达，差异有统计学意义（Z分别为－6.70和－4.09，P均〈0.05）；AML初发组LC3Ⅱ的表达量与健康对照组，缓解组和难治组相比均高表达，差异有统计学意义（Z分别为－6.96、－5.32和－3.52，P均〈0.05）；LC3Ⅱ高表达组患者细胞遗传学异常更为常见（χ2＝6.510，P〈0.ObjectiveTodetect the mechanism of acute myeloid leukemia（AML） and the relationship between autophagy andubiquitin c-terminal hydrolases L3（ UCH-L3） for providing new targets and guiding clinical management in AML.Methods84 cases of AML patients and 25 controls were chosen from the First Affiliated Hospital of Dalian Medical University from 2014 to 2015, including 47 males and 37 females. The AML patients were divided into 3 groups ： initialdiagnosis group[40 cases including 24 males and 16 females, with an median age of 54 （23-85）]; complete remission group[30 cases including 14 males and 16 females, with an median age of 45 （22-74）]and refractory group[14 cases including 9 males and 5 females, with an median age of 50 （18-80）]. Among those patients, there were 3 cases of M1, 42 cases of M2, 18 cases of M3, 3 cases of M4 and 18 cases of M5 subtypes.The expression of UCH-L3 and LC3II were detected by Real Time PCR and Western blot after the HL-60 cells were treated by TCID. The expression of UCH-L3 and LC3IIin the PBMC of AML patients and controls were detected with Real Time PCR. The expression levels of UCH-L3 and LC3II in different types of AML were analyzed. The relationship between the expression of LC3Ⅱand clinical features, clinical stages, laboratory results and therapeutic effects of patients were investigated. It further detected the expression of UCH-L3 and LC3IImRNA in post-induction status. t test were used for measurement data, χ2 test were performed for rate comparison; rank correlation test were performed for correlation analysis.Non-parametric test were performed for non-normal distribution data.ResultsBoth the UCH-L3 and LC3 II were down-regulated after TCID treatment in HL-60 cellsat gene and protein levels （t=-29.435, t=-8.105, P〈0.05）. The expression of UCH-L3 in initial diagnosis group was lower than control group（Z=-3.87, P〈0.05）, butit was higher in remission group than initial diagnosis group and refractory group（Z=-6.70 , Z=-4.09, P〈0.05）. The e

关 键 词：白血病 髓样 急性 半胱氨酸内肽酶类 微管相关蛋白质类 自噬 

分 类 号：R730.43[医药卫生—肿瘤] R733.71[医药卫生—临床医学]