骨髓增殖性疾病患者JAK2基因V617F突变频率、磷酸化JAK/STAT蛋白表达水平及临床意义

目的 探讨145例骨髓增殖件疾病患者JAK2基因V617F突变率;并分析JAK2基因V617F突变阳性患者的临床特点及意义.方法 应用PCR产物直接测序和等位基因特异性PCR技术检测145例骨髓增殖性疾病患者JAK2基因V617F突变.并应用Western blot方法测定JAK2基因V617F突变阳性患者JAK2蛋白、磷酸化JAK2蛋白及磷酸化STAT5蛋白表达;对JAK2基因V617F突变阳性与突变阴性的骨髓增殖性疾病患者临床资料进行比较,评价JAK2基因V617F突变阳性的临床意义.结果 ①本组病例真性红细胞增多症(PV)、特发性骨髓纤维化(ET)、原发性血小板增多症(IMF)患者JAK2基因V617F突变率分别为92%(64例中59例)、58%(43例中25例)和50%(38例中19例),等位基因特异性PCR检测较PCR产物直接测序有更高的JAK2基因V617F检出率PV84%(64例中53例)、MIF 44%(43例中19例)、ET 39%(38例中15例)].②JAK2基因V617F突变阳性患者外周血单个核细胞磷酸化JAK2及磷酸化STAT5蛋白表达较突变阴性者明显增高(P<0.05).③JAK2基因V617F突变阳性骨髓增殖性疾病患者发病平均年龄偏大;平均白细胞计数高于突变阴性患者;血小板计数小于突变阴性患者;脾脏较突变阴性患者小;JAK2基因V617F突变阳性PV、ET和IMF患者血栓性事什发生率分别为17%、32%和16%;而突变阴性PV、ET和IMF患者血栓性事件发生率分别为0、16%和5%.结论 骨髓增殖性疾病患者有较高的JAK2基因V617F突变发生率,且JAK2基因V617F突变阳性患者易发生血栓事件.

The frequency of JAK2 V617F mutation, expression level of phosphorylated JAK/STATs proteins and their clinical significance in myeloproliferative disorders patients

Objective To investigate the frequency of JAK2 V617F mutation in 145 myeloprolifera-tive disorders(MPDs) patients, analyze the correlation between JAK2 V617F mutation and clinical features. Methods The JAK2 V617F mutation was detected by direct DNA sequencing of PCR product and allele-spe-cific PCR respectively. The expression of JAK2, phospho-JAK2 and phuspho-STAT5 proteins was determined by Western blot. The clinical data of MPDs patients with or without JAK2 V617F mutation was collected and JAK2 V617F mutation for PV,IMF,ET was 92% ,58% ,50 % respectively. Compared with conventional DNA sequencing (PV 84%, IMF 44%, ET 39%, respectively), allele-specific PCR exhibited a higher sensitivity eral blood mononuclear cells (PBMNCs) were uprcgulated significantly in JAK2 V617F-positive patients than 617F-positive patients' features were as follows: older age of onset,higher mean leukocyte counts, lower platelet counts and smaller spleen volume. Frequency of thrombosis events in PT, ET, IMF was 17%, 32%, 16% respectively for JAK2 V617F positive group, and 0% (PV), 16% (ET) ,5% (IMF) for JAK2 V617F negative group. Conclusions MPDs patients display higher frequency of JAK2 V617F mutation. JAK2 V617F mutation positive patients predispose to a thrombosis tendency.