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Wilms基因(WT-1)最早于1990年由CALL等在Wilms瘤中发现,定位于人类染色体11p13[1],最早发现与肾母细胞瘤的发病机制有关,命名为Wilms瘤Ⅰ型基因[2]。研究[3-4]发现,WT-1可调控造血细胞增生和分化基因的转录或激活,与急性髓系白血病(AML)等多种肿瘤性疾病密切相关。因此,可作为白血病的特异性分子标志。本研究通过测定WT-1基因在非肿瘤病人和AML病人骨髓细胞的表达水平,比较AML病人和对照组以及AML各亚型WT-1基因水平的差异,并分析其与病人临床特征、疗效及预后的关系,为应用WT-1监测AML病人微小残留病和对AML病人进行预后分层提供理论依据。
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66例AML病人,59例检测到WT-1的表达,阳性率为89.4%;20例非肿瘤病人,6例检测WT-1的表达,阳性率为30%,2组WT-1表达率相比差异有统计学意义(χ2=29.34,P<0.01)。各组WT-1表达水平相比差异有统计学意义(F=7.206,P<0.05),与对照组相比,APL病人和ANLL中分组为M1的病人的WT-1表达水平差异均有统计学意义(P<0.05)。ANLL病人各亚型间相比差异有统计学意义(F=4.981,P<0.05),与M1组病人相比,M2组和M3组病人的WT-1表达水平差异有统计学意义(P<0.05)(见表 1)。
分组 n 表达水平 APL 8 0.600 0±0.601 5* ANLL M1 9 0.930 0±0.846 1* M2 29 0.297 2±0.341 4△ M4 7 0.198 4±0.226 3△ M5 2 0.153 4±0.166 4 对照组 20 0.000 2±0.000 3 F — 7.21 P — <0.01 MS组内 — 0.172 与对照组比较*P<0.05;与M1组比较△P<0.05 表 1 不同组间WT-1转录本表达水平比较(x±s)
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以58例ANLL病人WT-1的中位表达水平作为界值,将其分为WT-1高表达组和低表达组,不同WT-1表达组AML病人性别、年龄、骨髓原幼细胞比例、外周血白细胞数、血红蛋白、血小板数、乳酸脱氢酶(LDH)、C反应蛋白(CRP)、降钙素原(PCT)等临床指标统计见表 2。结果显示:与WT-1低表达组相比,WT-1高表达组中骨髓原幼细胞比例、外周血白细胞数、CRP显著升高(P<0.05);而性别、年龄、血红蛋白、血小板数、LDH、PCT差异无统计学意义(P>0.05)。
分组 男 女 年龄/岁 原幼细胞比例/% 外周血白细胞数/(×109/L) 血红蛋白/(g/L) 血小板数/(×109/L) LDH/(U/L) CRP/(mg/L) PCT/(ng/L) WT-1低表达组 14 15 51.1±15.8 45.69±21.16 13.80±17.83 90.00±27.72 58.79±67.76 493.97±592.42 35.20±30.89 0.35±0.23 WT-1高表达组 16 13 51.60±15.7 72.97±19.01 70.01±97.75 87.31±31.19 44.10±34.15 707.72±767.84 61.20±31.02 0.42±0.26 t 0.15* 0.12 5.16 3.05 0.35 1.04 1.19 3.71 1.09 P >0.05 >0.05 <0.05 <0.05 >0.05 >0.05 >0.05 < 0.05 >0.05 *示χ2值 表 2 WT1的表达与初诊ANLL病人临床特征比较(x±s)
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58例ANLL病人完善染色体核型分析及基因检查,WT-1高表达组有10例存在染色体核型异常,其中t(8;21)(q22;q22)/AML1-ETO和11p+各2例,inv(16)(p13q22)/CBFB-MYH11、-y、8q-、+11、+21、del(20)(q11, 2q3.1)各1例,基因突变检测到NPM1突变2例,C/EBPα突变1例,C-kit突变1例,FLT3-ITD突变1例,NPM1、FLT3-ITD、DNMT3α突变1例;WT-1低表达组有6例染色核型异常,其中t (8;21)(q22;q22)、inv(16) (p13q22)、del(12)(q22q35)、del(5)(p11, 2p13)、多倍体、复杂核型各1例,C-kit突变1例,NPM1突变1例,C/EBPα突变2例,FLT3-ITD突变1例,NPM1、FLT3-TKD突变2例,NPM1、DNMT3α突变1例。根据《成人急性髓系白血病(非急性早幼粒细胞白血病)中国诊疗指南(2017版)》进行预后危险度分级,WT-1高表达组预后良好4例,预后中等22例,预后不良3例,WT-1低表达组预后良好6例,预后中等20例,预后不良3例,2组间病人预后水平差异无统计学意义(uc=394.50, P>0.05)(见表 3)。
分组 初次缓解 初次未缓解 半年复发 半年未复发 WT-1高表达组 16 13 8 21 WT-1低表达组 24 5 4 25 χ2 3.95 0.95 P <0.05 >0.05 表 3 不同WT-1表达组AML病人初次诱导后疗效评估(ni=29)
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8例APL病人在观察期内,2例病人发生颅内出血死亡,其余6例均初次诱导达完全缓解(CR)。58例ANLL病人中,WT-1高表达组与低表达组初次诱导CR率分别为53.6 %和82.8 %,高表达组AML病人CR率显著低于低表达组(P<0.05);WT-1高表达组与低表达组获得CR的病人半年内复发率分别为29.2%和14.3%,2组差异无统计学意义(P>0.05)(见表 3)。
定量检测WT-1基因在急性白血病病人中的表达及临床意义
Quantitative detection of the WT-1 gene expression in acute leukemia patients and its clinical significance
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摘要:
目的研究Wilms基因(WT-1)在急性髓系白血病(AML)病人骨髓细胞中的表达,并分析其表达与临床疗效的关系。 方法收集AML病人66例,其中急性早幼粒细胞白血病(APL)组8例,急性非淋巴细胞白血病(ANLL,除APL)组58例。对照组20名为健康体检者。采用实时定量PCR法检测骨髓细胞中WT-1表达,并以WT-1的中位表达水平作为界值,将初诊AML病人(APL除外)分为WT-1高表达组和低表达组,并分析其表达与病人性别、年龄、骨髓原幼细胞比例、外周血白细胞数、血红蛋白、血小板数、乳酸脱氢酶(LDH)、C反应蛋白(CRP)、降钙素原(PCT)、完全缓解(CR)率及复发率的关系,以揭示WT-1基因表达水平与AML预后的关系。 结果病例组与对照组相比WT-1表达率差异有统计学意义(χ2=29.34,P < 0.01)。各组WT-1表达水平相比差异有统计学意义(F=7.206,P < 0.05),与对照组相比,APL病人和ANLL中M1的病人的WT-1表达水平均较高(P < 0.05)。ANLL病人各亚型间相比差异有统计学意义(F=4.981,P < 0.05),与M1组病人相比,M2组和M3组病人的WT-1表达水平相对较低(P < 0.05);与WT-1低表达组相比,WT-1高表达组中骨髓原幼细胞比例、外周血白细胞数显著升高(P < 0.05);而性别、年龄、血红蛋白、血小板数、LDH、CRP、PCT差异无统计学意义(P>0.05),2组病人预后水平差异无统计学意义(P>0.05);AML病人WT-1高表达组CR率低于低表达组(P < 0.05)。 结论WT-1在AML病人骨髓细胞中表达水平可作为临床评估疾病的预后、疗效的指标。 Abstract:ObjectiveTo study the expression of Wilms gene (WT-1) in bone marrow cells of patients with acute myeloid leukemia (AML), and analyze the relationship between the expression level of WT-1 and clinical efficacy. MethodsThe clinical data of 66 AML patients, which included 8 patients with acute promyelocytic leukemia (APL) and 58 patients with acute non-lymphocytic leukemia (ANLL, except APL), were collected.Twenty healthy people were set as the control group.The expression levels of WT-1 in bone marrow cells of AML patients were detected using real-time quantitative PCR.The median level of WT-1 was set as the clinic value, the 58 newly diagnosed as ANLL patients were divided into the high-expression WT-1 group and low-expression WT-1 group.The relationship between the expression level of WT-1 and sex, age, ratio of progenitor cells, count of peripheral white blood cells, hemoglobin, platelets, lactate dehydrogenase (LDH), C-reactive protein (CRP), calcitonin (PCT), complete remission rate (CR) and relapse rate were analyzed, and the relationship between WT-1 gene expression level and prognosis of AML patients was revealed. ResultsThe difference of the expression rate of WT-1 between cases group and control group was statistically significant (P < 0.01), and the difference of the expression level of WT-1 among each group was statistically significant (P < 0.05).Compared with the control group, the expression levels of WT-1 in APL patients and M1 group were higher (P < 0.05).The difference of the expression level of WT-1 among ANLL patient subtypes was statistically significant (P < 0.05).Compared with the M1 group, the expression levels of WT-1 in M2 and M3 group were relatively low (P < 0.05).Compared with the low WT-1 expression group, the proportion of bone marrow progenitor cells and number of peripheral blood white blood cells in high WT-1 expression group significantly increased (P < 0.05).There was no statistical significance in gender, age, hemoglobin, platelet count, LDH, CRP, and PCT between low and high WT-1 expression groups (P>0.05), and there was no statistical significance in prognosis between two groups (P>0.05).The CR rate of AML patients in high WT-1 expression group was lower than that in low WT-1 expression group (P < 0.05). ConclusionsThe expression level of WT-1 in bone marrow cells of patients with AML can be used as an index to evaluate the prognosis and curative effect of the disease. -
Key words:
- acute myeloid leukaemia /
- WT-1 gene /
- real-time quantitative PCR
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表 1 不同组间WT-1转录本表达水平比较(x±s)
分组 n 表达水平 APL 8 0.600 0±0.601 5* ANLL M1 9 0.930 0±0.846 1* M2 29 0.297 2±0.341 4△ M4 7 0.198 4±0.226 3△ M5 2 0.153 4±0.166 4 对照组 20 0.000 2±0.000 3 F — 7.21 P — <0.01 MS组内 — 0.172 与对照组比较*P<0.05;与M1组比较△P<0.05 表 2 WT1的表达与初诊ANLL病人临床特征比较(x±s)
分组 男 女 年龄/岁 原幼细胞比例/% 外周血白细胞数/(×109/L) 血红蛋白/(g/L) 血小板数/(×109/L) LDH/(U/L) CRP/(mg/L) PCT/(ng/L) WT-1低表达组 14 15 51.1±15.8 45.69±21.16 13.80±17.83 90.00±27.72 58.79±67.76 493.97±592.42 35.20±30.89 0.35±0.23 WT-1高表达组 16 13 51.60±15.7 72.97±19.01 70.01±97.75 87.31±31.19 44.10±34.15 707.72±767.84 61.20±31.02 0.42±0.26 t 0.15* 0.12 5.16 3.05 0.35 1.04 1.19 3.71 1.09 P >0.05 >0.05 <0.05 <0.05 >0.05 >0.05 >0.05 < 0.05 >0.05 *示χ2值 表 3 不同WT-1表达组AML病人初次诱导后疗效评估(ni=29)
分组 初次缓解 初次未缓解 半年复发 半年未复发 WT-1高表达组 16 13 8 21 WT-1低表达组 24 5 4 25 χ2 3.95 0.95 P <0.05 >0.05 -
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