血浆游离DNA检测FLT3-ITD突变及ITD特征对急性髓细胞白血病诊断的意义  

Significance of FLT3-ITD mutation and the characteristics of ITD in plasma free DNA in the diagnosis of acute myeloid leukemia

作　　者：王楠[1] 吴丹[2] 赵乙洁[1] 赵婧媛[3] 姜凤全 袁宏[1] 

机构地区：[1]大连医科大学附属第一医院检验科,辽宁大连116023 [2]大连医科大学附属第二医院检验科,辽宁大连116011 [3]大连医科大学附属第一医院再生医学研究中心,辽宁大连116023

出　　处：《临床检验杂志》2015年第2期111-114,118,共5页Chinese Journal of Clinical Laboratory Science

基　　金：辽宁省科技厅项目(20102047)

摘　　要：目的探讨在血浆游离DNA中检测Fms样酪氨酸激酶3(FLT3)基因内部串联重复序列(ITD)突变及ITD特征,为急性髓细胞白血病(AML)无创性诊断、个体化的分子靶向治疗提供新的策略。方法提取88例初诊AML患者和40例对照者(体检健康者和血液系统良性疾病患者)的血浆游离DNA及骨髓细胞DNA并检测其浓度;PCR检测血浆及骨髓DNA中FLT3-ITD突变的表达水平;毛细管电泳法检测FLT3-ITD阳性患者血浆游离DNA和骨髓细胞DNA中的ITD数量,并进行统计学分析。结果初诊AML患者组血浆ccf DNA浓度为203.85(115.8,1 165.7)μg/L,而体检健康者和血液系统良性疾病患者分别为4.60(3.21,5.79)μg/L和9.26(5.23,16.77)μg/L,三组间差异有统计学意义(K-Wχ2=84.64,P<0.05);初诊AML患者组血浆ccf DNA浓度显著高于血液系统良性病变患者组及体检健康组(Z分别为-7.10和-6.96,P均<0.05),而体检健康者与血液系统良性疾病患者间、FAB分型不同的初诊AML患者间的血浆游离DNA浓度差异无统计学意义(Z=-1.56,P>0.05;F=0.073,P>0.05);FAB各型别中M2型FLT3-ITD突变率最高为30.77%(12/39),其次为M4型(20%,1/5),M5型(20%,2/10)和M3型(12%,3/25),其余型别未见突变;发生FLT3-ITD突变18例,与骨髓细胞DNA检测结果一致,13例患者在毛细管电泳中出现一条ITD条带峰,骨髓细胞结果与血浆结果一致;5例出现多条ITD条带峰,其骨髓细胞结果只有2例一致。结论监测血浆游离DNA中FLT3-ITD突变及ITD数量,可能会成为AML个体化诊断的新分子标志物。Objective To investigate the mutation of Fms-like tyrosine kinase-3（ FLT3）-internal tandem duplication（ ITD） and the characteristics of ITD in plasma free DNA,and provide a new strategy for the noninvasive diagnosis and individualized molecular targeted therapy of acute myeloid leukemia（ AML）. Methods Plasma free DNA and bone marrow cell DNA samples from 88 patients with prelimilary diagnosis of AML,20 patients with benign hematological system diseases and 20 healthy controls were obtained,and the concentration of DNA was determined. The mutations of FLT3-ITD in these DNA samples were detected by PCR. The number of ITD in the patients with FLT3-ITD mutation was detected by the capillary electrophoresis. The obtained results were analyzed and compared.Results The concentration（ median [P25,P75]） of plasma free DNA in the patients with preliminary diagnosis of AML was 203. 85（ 115. 8,1 165. 7） μg/L,while those in the patients with benign hematological system diseases and the healthy control were 9. 26（ 5. 23,16. 77） μg/L and 4. 60（ 3. 21,5. 79） μg/L,respectively. There was significant difference among the three groups（ K-Wχ^2= 84. 64,P〈0. 05）,and the former was significantly higher than the latter two（ Z =- 7. 10 and- 6. 96,P〈0. 05）,but there was no significant difference between the latter two（ Z =- 1. 56,P〉0. 05）. Moreover,there was no significant difference in the concentration of plasma free DNA between different FAB typings of AML（ K-W χ^2= 3. 34,P〉0. 05）. The mutation rate of FLT3-ITD was up to 30. 77%（ 12 /39） in FAB-M2,followed by FAB-M4（ 20%,1 /5）,M5（ 20%,2 /10） and M3（ 12%,3 /25）,and zero for the rest types of FAB. A total of 18 patients were detected the mutation of FLT3-ITD,and there was consistent results between plasma DNA and bone marrow DNA. Among them,13 were detected one band of ITD in the capillary electrophoresis,and 5 with multiple bands. There was consistent results in the former between plasma DNA and bone marrow DNA,whi

关 键 词：急性髓细胞白血病 血浆游离DNA Fms样酪氨酸激酶3-串联重复序列 

分 类 号：R733.71[医药卫生—肿瘤]