6例急性巨核细胞白血病患儿实验室诊断分析

doi:10.3969/j.issn.1673-8640.2025.05.012

摘要/Abstract

摘要：

目的分析急性巨核细胞白血病（AMKL）患儿的实验室诊断方法。方法选取2020年1月—2023年12月上海市儿童医院6例AMKL患儿，进行骨髓细胞形态学FAB分型、免疫表型、白血病细胞表面相关抗原、骨髓细胞24 h培养G显带染色体核型、白血病常见融合基因和单个基因表达、血液肿瘤突变基因和全转录组检测。结果 6例AMKL患儿的骨髓原始巨核细胞比例均≥20%；糖原染色（PAS）、α-醋酸萘酚酯酶染色（α-NAE）均为阳性，过氧化物酶染色（POX）和氟化钠抑制试验均为阴性；6例患儿均表达cCD41，有5例表达CD61；有3例患儿染色体核型正常、2例核型为复杂核型、1例核型有12号染色体结构异常；有5例患儿WT1基因阳性；2例行血液肿瘤全转录组检测的患儿中，1例NUP98-KDM5A融合基因阳性、1例CBFA2T3-GLIS2融合基因阳性。结论儿童AMKL的实验室诊断应联合骨髓细胞形态学、流式细胞免疫分型、细胞遗传学和分子生物学等方法检测结果综合分析，以有效指导临床制定治疗方案，准确判断疗效和预后。

关键词: 急性巨核细胞白血病, 细胞形态学, 免疫学, 细胞遗传学, 分子生物学分型, 二代测序, 儿童

Abstract:

Objective To analyze the laboratory diagnostic methods of acute megakaryocytic leukemia （AMKL） in children. Methods Totally，6 cases of AMKL from Shanghai Children's Hospital from January 2020 to December 2023 were enrolled for morphological FAB typing of bone marrow cells，immunophenotyping，surface associated antigen of leukemia cells，24 h culture G-banding chromosome karyotype of bone marrow cells，expressions of common fusion genes and single genes of leukemia，blood tumor mutation genes and whole transcriptomes. Results The proportion of bone marrow primitive megakaryocytes in 6 cases of AMKL was≥20%. Periodic acid-schiff stain （PAS） and alpha-naphthol acetate esterase staining （α-NAE） were both positive，while peroxidase stain （POX） and sodium fluoride inhibition test were both negative. The 6 cases all expressed cCD41，and 5 cases were CD61. Totally，3 cases had normal chromosome karyotypes，2 cases had complex chromosome karyotypes，and 1 case had structural abnormalities on chromosome 12. Five cases had WT1 gene positivity. One case of NUP98-KDM5A fusion gene was positive，and another case of CBFA2T3-GLIS2 fusion gene was positive in 2 routine blood tumor transcriptome test cases. Conclusions It is necessary to comprehensively analyze and diagnose AMKL in children by combining bone marrow cell morphology，flow cytometry immunophenotyping，cytogenetics and molecular biology determination methods，which is of significance for guiding the selection of clinical treatment plans and efficacy and prognosis judgment.

Key words: Acute megakaryocytic leukemia, Morphology, Immunology, Cytogenetics, Molecular biology, Next generation sequencing, Children

中图分类号:

R733.71

柳敏, 孙恒娟, 杜成坎, 夏敏, 姜林林, 翁文浩, 张泓. 6例急性巨核细胞白血病患儿实验室诊断分析[J]. 检验医学, 2025, 40(5): 484-488.

LIU Min, SUN Hengjuan, DU Chengkan, XIA Min, JIANG Linlin, WENG Wenhao, ZHANG Hong. Laboratory analysis of 6 cases of acute megakaryocytic leukemia in children[J]. Laboratory Medicine, 2025, 40(5): 484-488.

图/表 5

参考文献 20

[1]	HAMA A, YAGASAKI H, TAKAHASHI Y, et al. Acute megakaryoblastic leukaemia(AMKL)in children:a comparison of AMKL with and without Down syndrome[J]. Br J Haematol, 2008, 140(5):552-561.
[2]	DOWLING G P, PICCIN A, GAVIN K T, et al. A retrospective study of myeloid leukaemia in children with Down syndrome in Ireland[J]. Ir J Med Sci, 2020, 189(3):979-984.
[3]	WANG Y, LU A, JIA Y, et al. Outcome and prognostic features in pediatric acute megakaryoblastic leukemia without Down syndrome:a retrospective study in China[J]. Clin Lymphoma Myeloma Leuk, 2021, 21(4):e301-e308.
[4]	ARBER D A, ORAZI A, HASSERJIAN R, et al. The 2016 revision to the world health organization classification of myeloid neoplasms and acute leukemia[J]. Blood. 2016, 127(20):2391-2405. DOI PMID
[5]	MCGOWAN-JORDAN J, SIMONS A, SCHMID M. ISCN 2016:an international system for human cytogenomic nomenclature(2016)[M]. Basel:S. Karger AG, 2016.
[6]	BALANDRÁN J C, VADILLO E, DOZAL D, et al. Analysis of normal hematopoietic stem and progenitor cell contents in childhood acute leukemia bone marrow[J]. Arch Med Res, 2016, 47(8):629-643. DOI PMID
[7]	O'BRIEN M M, CAO X, POUNDS S, et al. Prognostic features in acute megakaryoblastic leukemia in children without Down syndrome:a report from the AML02 multicenter trial and the Children's Oncology Group Study POG 9421[J]. Leukemia, 2013, 27(3):731-734.
[8]	MARQUES-PIUBELLI M L, CORDEIRO M G, CRISTOFANI L, et al. Acute megakaryoblastic leukemia with t(1;22)(p13.3;q13.1);RBM15-MKL1 mimicking hepatoblastoma in an infant:the role of karyotype in differential diagnosis[J]. Pediatr Blood Cancer, 2020, 67(3):e28111.
[9]	刘彦权, 殷悦, 陈玉婷, 等. 急性巨核细胞白血病14例临床分析并文献复习[J]. 解放军医学杂志, 2022, 47(10):1006-1012.
[10]	CHETTY T, MASINGI NI, LAHER Z, et al. Acute megakaryoblastic leukaemia:light microscopy and scanning electron microscopy of blast cells[J]. Br J Haematol, 2017, 176(5):686.
[11]	TALLMAN M S, NEUBERG D, BENNETT J M, et al. Acute megakaryocytic leukemia:the Eastern Cooperative Oncology Group experience[J]. Blood, 2000, 96(7):2405-2411.
[12]	蓝建平, 孙酬经, 钱美华, 等. 急性巨核细胞白血病的免疫表型分析[J]. 浙江临床医学, 2000, 2(7):441-443.
[13]	EIDENSCHINK BRODERSEN L, ALONZO T A, MENSSEN A J, et al. A recurrent immunophenotype at diagnosis independently identifies high-risk pediatric acute myeloid leukemia:a report from Children's Oncology Group[J]. Leukemia, 2016, 30(10):2077-2080.
[14]	ZHANG W, DUN J, LI H, et al. Analysis 33 patients of non-DS-AMKL with or without acquired trisomy 21 from multiple centers and compared to 118 AML patients[J]. Hematology, 2023, 28(1):2231731.
[15]	DE ROOIJ J D, MASETTI R, VAN DEN HEUVEL-EIBRINK M M, et al. Recurrent abnormalities can be used for risk group stratification in pediatric AMKL:a retrospective intergroup study[J]. Blood, 2016, 127(26):3424-3430.
[16]	PASTORE F, GITTINGER H, RAAB S, et al. Acute megakaryoblastic leukaemia shows high frequency of chromosome 1q aberrations and dismal outcome[J]. Br J Haematol, 2023, 202(6):1165-1177.
[17]	罗铁梅, 于洁, 安曦洲. 儿童急性巨核细胞白血病临床特点及预后分析[J]. 中国当代儿科杂志, 2021, 23(6):613-620.
[18]	DE ROOIJ J D, BRANSTETTER C, MA J, et al. Pediatric non-Down syndrome acute megakaryoblastic leukemia is characterized by distinct genomic subsets with varying outcomes[J]. Nat Genet, 2017, 49(3):451-456. DOI PMID
[19]	HARA Y, SHIBA N, OHKI K, et al. Prognostic impact of specific molecular profiles in pediatric acute megakaryoblastic leukemia in non-Down syndrome[J]. Genes Chromosomes Cancer, 2017, 56(5):394-404.
[20]	ZANGRANDO A, CAVAGNERO F, SCARPARO P, et al. CD56,HLA-DR,and CD45 recognize a subtype of childhood AML harboring CBFA2T3-GLIS2 fusion transcript[J]. Cytometry A, 2021, 99(8):844-850.

病例编号	性别	年龄/月	血象				骨髓象		临床体征
病例编号	性别	年龄/月	白细胞/ （×109L^-1）	血红蛋白/ （g·L^-1）	血小板/ （×109L^-1）	原始细胞/%	增生程度	原始细胞/ %	贫血	出血	发热	肝肿大	脾肿大
1	女	18.0	18.10	98	18	29	明显活跃	28.5	有	有	有	1 cm	无
2	男	7.6	7.27	91	12	0	活跃	24.0	有	无	有	1 cm	5 cm
3	女	82.0	8.58	106	59	0	明显活跃	88.5	有	无	有	5 cm	无
4	男	58.0	8.48	120	67	0	明显活跃	23.5	无	有	无	无	无
5	男	9.7	13.39	84	20	0	活跃	20.0	有	有	无	无	无
6	男	17.0	8.30	93	105	0	活跃	91.0	有	无	有	4 cm	1 cm

病例编号	性别	年龄/月	血象				骨髓象		临床体征
病例编号	性别	年龄/月	白细胞/ （×109L^-1）	血红蛋白/ （g·L^-1）	血小板/ （×109L^-1）	原始细胞/%	增生程度	原始细胞/ %	贫血	出血	发热	肝肿大	脾肿大
1	女	18.0	18.10	98	18	29	明显活跃	28.5	有	有	有	1 cm	无
2	男	7.6	7.27	91	12	0	活跃	24.0	有	无	有	1 cm	5 cm
3	女	82.0	8.58	106	59	0	明显活跃	88.5	有	无	有	5 cm	无
4	男	58.0	8.48	120	67	0	明显活跃	23.5	无	有	无	无	无
5	男	9.7	13.39	84	20	0	活跃	20.0	有	有	无	无	无
6	男	17.0	8.30	93	105	0	活跃	91.0	有	无	有	4 cm	1 cm

病例编号	CD13	CD33	CD34	CD36	cCD41	CD56	CD61	CD117	HLA-DR
1	0	89.6	0	91.8	73.6	0	28.2	0	86.3
2	0	80.8	17.1	0	57.2	0	37.0	63.1	9.8
3	38.7	32.7	43.9	0	86.6	0	69.8	0	0
4	27.8	87.6	54.2	90.7	87.9	0	89.9	66.7	56.1
5	27.0	78.6	0	83.6	37.3	0	36.0	0	30.5
6	0	97.2	78.2	0	16.6	98.0	0	93.8	0

病例编号	CD13	CD33	CD34	CD36	cCD41	CD56	CD61	CD117	HLA-DR
1	0	89.6	0	91.8	73.6	0	28.2	0	86.3
2	0	80.8	17.1	0	57.2	0	37.0	63.1	9.8
3	38.7	32.7	43.9	0	86.6	0	69.8	0	0
4	27.8	87.6	54.2	90.7	87.9	0	89.9	66.7	56.1
5	27.0	78.6	0	83.6	37.3	0	36.0	0	30.5
6	0	97.2	78.2	0	16.6	98.0	0	93.8	0

病例编号	染色体核型分析	qPCR	二代测序
病例编号	染色体核型分析	常见融合基因和单个基因	血液肿瘤突变基因	血液肿瘤全转录组
1	50~51，XX，add（3）（q21），?del（4）（q22）， ?+19，+mar1~3，inc[cp2]/46，XX[8]	WT1阳性	未检测到突变	未送检
2	46，XY[6]	WT1阳性	TET2突变型	未送检
3	46，XX[5]	WT1阳性	未检测到突变	未送检
4	46，XY[20]	WT1阳性	未检测到突变	未送检
5	46，XY，inv（12）（p13q21）[3]/46，XY[17]	阴性	未检测到突变	NUP98-KDM5A阳性
6	46，X，-Y，+3，add（16）（q12-13）[4]/47， idem，+mar[4]/46，XY[12]	WT1阳性	未检测到突变	CBFA2T3-GLIS2阳性