骨髓增生异常综合征ASXL1基因突变的研究  

Mutations of ASXLI gene in myelodysplastic syndromes

作　　者：刘海霞[1] 王宏伟[1] 覃艳红[1] 陈秀花[1] 任方刚[1] 常建梅[1] 张耀方[1] 薛凤[1] 李娟[1] 徐智芳[1] Liu Haixia Wang Hongwei Tan Yanhong Chen Xiuhua Ren Fanggang Chang Jianmei Zhang Yaofang Xue Feng Li Juan Xu Zhifang(Institute of Hematology, the Second Hospital of Shanxi Medical University, Shanxi Key Laboratory of Molecular Diagnosis and Treatment of Blood Diseases, Taiyuan 030001, China)

机构地区：[1]山西医科大学第二医院血液病研究所血液病分子诊疗山西省重点实验室,太原030001

出　　处：《白血病．淋巴瘤》2017年第9期513-518,共6页Journal of Leukemia & Lymphoma

基　　金：国家自然科学基金（81200390）;中国博士后科学基金（20100471583）

摘　　要：目的研究骨髓增生异常综合征（MDS）患者中表观遗传调节因子ASXL1基因的突变情况。方法在DNA水平采用聚合酶链反应（PCR）扩增产物片段直接测序分析法检测53例初发MDS患者及20名健康人ASXL1基因第12外显子突变情况，比较ASXL1突变患者与野生型患者临床及实验室特征；反转录聚合酶链反应（RT-PCR）扩增产物直接测序分析法检测ASXL1突变在mRNA水平的表达。结果在53例MDS患者中，9例（16.9 %）ASXL1基因突变。共检测出6种突变类型，包括4种移码突变（2例p.Glu635ArgfsX15、3例p.Gly646TrpfsX12、1例p.Ala640GlyfsX14、1例p.Gly790TrpfsX10）和2种无义突变（1例p.Gln1063X和1例p.Gln695X）。所有突变类型均为杂合突变，其中p.Gly790TrpfsX10和p.Gln695X为新发现突变类型。此外还检测到一种单核苷酸多态性（SNP）位点（4例p.Gly652Ser）。20名健康人中检测出pGly652Ser SNP 5名和p.Leu1173Leu SNP 1名。RT-PCR扩增产物片段直接测序可在mRNA水平检测出移码突变（p.Gly646TrpfsX12）。ASXL1突变患者初发时外周血白细胞计数、红细胞计数、血小板计数、血红蛋白、网织红细胞、中性粒细胞绝对值、外周血淋巴细胞比例、T细胞亚群、骨髓原始细胞比例、MDS分型和染色体核型分布与ASXL1野生型患者相比，差异均无统计学意义（均P〉0.05）。结论ASXL1基因在MDS患者中的突变频率较高，且可在mRNA水平检测到ASXL1基因突变。Objective To investigate the mutations of epigenetic regulation factor ASXL1 gene in myelodysplastic syndrome （MDS）. Methods Mutation analysis of ASXL1 gene in 53 de novo MDS patients and 20 healthy persons was performed by using polymerase chain reaction （PCR） followed by sequence analysis at DNA level. The clinical and laboratory characteristics were compared in MDS patients with ASXL1 gene mutation and ASXL1 wild type. ASXL1 mutation in mRNA level was detected by using reverse transcription PCR （RT-PCR） followed by sequence analysis. Results ASXL1 gene mutations were observed in 9 eases （16.9 %） of 53 MDS patients.6 mutation types were detected, including 4 frameshifl mutations types （2 cases with p.Glu635ArgfsX15, 3 cases with p.Gly646TrpfsX12, 1 case with p.Ala640GlyfsX14 and 1 case with p.Gly790TrpfsX10） and 2 nonsense mutation types （1 case with p.Gln1063X and 1 case with p.Gln695X）. All the mutations were heterozygous, and p.Gly790TrpfsX10 and p.Gln695X were new mutation types. In addition, a single nueletide polymorphism （SNP） p.Gly652Ser was also detected in 4 cases with MDS. 5 cases of p.G652S SNP and 1 case of p.Leu1173Leu SNP were detected in 20 healthy people. Frameshift mutation （p.Gly646TrpfsX12） could be detected at mRNA level by using RT-PCR. Differences were not observed in red blood cell counts, white blood cell counts, platelet counts, hemoglobin levels, reticulocyte, neutrophil granulocyte, the peripheral blood lymphocytes percentage, T-cell subsets in the peripheral blood, the proportion of primitive cell in the marrow and MDS types between the patients with ASXL1 gene mutation and ASXL1 wild type patients （P 〉 0.05）. Conclusion There is a high frequency of ASXL1 gene mutation in MDS patients, which can be detected at mRNA level.

关 键 词：基因 ASXL1 骨髓增生异常综合征 突变 表观遗传调节因子 

分 类 号：R551.3[医药卫生—血液循环系统疾病]