135例骨髓增殖性肿瘤患者JAK2基因突变的定量研究  

A quantitative assay for JAK2 mutation in 135 patients with chronic myeloproliferative neoplasms

作　　者：晁红颖[1] 沈益民[1] 张日[1] 冯宇峰[1] 岑建农[1] 姚利[1] 沈宏杰[1] 朱子玲[1] 薛永权[1] 

机构地区：[1]苏州大学附属第一医院、江苏省血液研究所,卫生部血栓与止血重点实验室,215006

出　　处：《中华血液学杂志》2009年第5期321-325,共5页Chinese Journal of Hematology

基　　金：江苏省卫生厅“科教兴卫工程”临床医学中心开放课题基金（WKF07006）

摘　　要：目的研究JAK2基因突变在骨髓增殖性肿瘤（MPN）中的发生率和突变类型，并对突变转录本水平进行定量分析，初步探讨其临床意义。方法采用突变序列扩增系统PCR（ARMS—PCR）法检测JAK2突变的发生率及其突变类型；采用毛细管电泳法定量分析JAK2突变转录本水平。结果135例MPN患者共检出95例JAK2V617F阳性，总阳性率为70．4％；真性红细胞增多症（PV）患者JAK2V617F突变发生率为97．4％，原发性血小板增多症（ET）为59．6％，3例特发性骨髓纤维化（IMF）中2例阳性；95例突变患者中纯合突变36例，杂合突变59例，其中PV患者纯合突变发生率为47．3％，高于ET患者的18．1％（P〈0．05）。纯合突变者其JAK2V617F突变转录本水平高于杂合突变者（P〈0．05），杂合型PV患者的JAK2V617F突变转录本水平高于杂合型ET患者（P〈0．05）；JAK2V617F突变高表达组（转录本水平为70％～100％）的平均年龄高于低表达组（转录本水平为47．3％～70．0％），且两者年龄均较JAK2V617F阴性组为高（P〈0．05）；年龄〈60岁者其JAK2V617F转录本水平低于≥60岁者（P〈0．001）；PV患者中，JAK2V617F高表达组其白细胞计数高于低表达组（P〈0．001），ET患者中，JAK2V617F高表达组其白细胞计数高于低表达组（P〈0．05），且二者均较阴性组为高（P〈0．05），血红蛋白水平在高表达组与低表达组间差异无统计学意义（P〉0．05），但二者均较阴性组为高（P〈0．05）。101例患者进行了染色体检查，未发现核型异常与JAK2V617F突变之间存在相关性。结论ARMS—PCR可作为检测JAK2V617F突变较灵敏的方法，结合毛细管电泳可用于此突变的定量分析以及临床MPN的诊断和微量残留病的检测。Objective To investigate the frequency and mutational status of JAK2 mutation in Chinese patients with chronic myeloproliferative neoplasms （MPN） and study the relative quantitation and clinical implications of mutated JAK2 transcript. Methods JAK2 mutation and the mutational status were screened with amplification-refractory mutation sequencing polymerase chain reaction（ARMS-PCR） , the relative quantity of mutated JAK2 mRNA by using capillary electrophoresis. Results JAK2V617F mutation was detected in 95 of 135 MPN patients, including 37 （97.4%） of 38 polycythemia vera（ PV）, 56 （59.6%） of 94 essential thrombocythemia （ET）and 2 of 3 idiopathic myelofibrosis （IMF）patients; the difference between the mutations in PV and ET was significant（P 〈 0.05 ）. Of 95 JAK2V617F patients examined, 18/38 PV patients （47.3%） and 17/94 （18.1%） ET patients and 1 IMF patient were homozygotes, and ET patients showed lower prevalence of homozygote （ P 〈 0.05 ）. In 95 MPN patients, the mutated mRNA ratio was higher in homozygotc than in heterozygote patients. PV heterozygote patients showed higher levels of mutated JAK2 mRNA than ET heterozygote patients（ P 〈 0.05 ）. The levels of JAK2V617F mRNA in patients over 60 years of age were significantly higher than that in those less than 60 years of age （P 〈 0. 001 ）. Higher leukocyte counts were observed in PV and ET patients with higher levels of mutated JAK2 mRNA （ P 〈 0.05 ）. The presence of JAK2V617F was found to be significantly associated with higher hemoglobin level in ET patients. Cytogeneticanalysis was performed in 101 of the 135 patients, the association between abnormal karyotype and JAK2V617F was not found. Conclusion The ARMS-PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation, and along with capillary electrophoresis, the estimation of minimal residual disease becomes possible.

关 键 词：骨髓增殖性疾病 DNA突变分析 突变序列扩增系统PCR 电泳 毛细管 

分 类 号：R686[医药卫生—骨科学]