急性白血病骨髓细胞线粒体DNA D-loop区的突变检测

摘要

本研究旨在检测急性白血病(AL)患者骨髓细胞线粒体DNA (mtDNA) D-loop区突变和线粒体微卫星不稳(mtMSI),并分析它们与AL发病的关系.选取19例初诊的AL患者,提取骨髓细胞mtDNA并对mtDNA D-loop区序列行PCR扩增,通过正反向直接测序对PCR扩增产物基因序列进行检测,并将测序结果与修订的剑桥标准序列(revised Cambridge reference sequence,rCRS)和有关数据库(MITOMAP数据库、GenBank数据库、mtDB数据库)进行比对.结果表明:AL的mtDNA D-loop区突变率达79％(15/19例),在D-loop区发现215个变异位点(35个突变位点、180个SNP位点)和1个mtMSI；发现1个新的突变类型nt150 C-CT,同时发现不同年龄、不同AL分型(AML,B-ALL)的患者间突变个数无统计学差异(P＞0.05).结论:AL的骨髓细胞mtDNA D-loop区存在高频率突变,且其突变可能与AL发病有关.%This study was aimed to detect the mutations and microsatellite instability (mtMSI) in mitochondrial DNA (mtDNA) D-loop region in bone marrow cells of acute leukemia (AL) patients, and to analyze their relationship with the pathogenesis of AL. 19 cases of newly diagnosed AL were enrolled in this study. Through extracting mtDNA, the D-loop region was amplified by polymerase chain reaction (PCR), the sequences of PCR products were detected by the pros- and cons-direct sequencing methods. The sequencing results were compared with the revised Cambridge reference sequence(rCRS) and the relevant database (MITOMAP database, GenBank database, mtDB database). The results showed that the mutation rate of mtDNA D-loop region in AL was 79% (15/19). 215 variations(35 mutations, 180 SNP) and a kind of mtMSI in the D-loop region were detected. A new type of mutation ntl50 C-CT was found. Also, there was no significant difference in the number of mutations between patients with different ages and different types of AL( AML, B-ALL). It is concluded that there is high frequency of mutations in the mtDNA D-loop, and the mutations may be associated with the pathogenesis of AL.