骨髓增殖性肿瘤中JAK2V617F突变和TNF-α的表达及两者的相关性研究

本研究旨在探讨骨髓增殖性肿瘤（MPN）患者外周血单个核细胞中JAK2V617F突变和TNF-α因子的表达情况及两者之间的相关性,为临床诊断及治疗提供理论依据.选取山东大学附属省立医院血液科确诊的62名BCR-ABL阴性的MPN患者（35例ET患者,19例PV患者,8例MF患者）及15例健康成年人为研究对象,将患者及正常对照者的外周血单个核细胞分为两部分,分别提取细胞中的DNA及mRNA,并将mRNA反转录为cDNA.采用SYBR Green Ⅰ实时荧光定量PCR分别检测JAK2V617F突变及TNF-α的表达量,并分析两者的相关性.结果表明,MPN患者JAK2V617F基因突变总体阳性率为64.52％ （40/62）,其中ET患者的JAK2V617F突变阳性率为54.28％ （19/35）,PV患者的JAK2V617F突变阳性率为94.74％（18/19）,MF患者的JAK2V617F突变阳性率为37.50％（3/8）;ET、PV、MF患者的JAK2V617F突变比例分别为0.838±0.419、4.417±0.658、2.746±2.009;ET、PV、MF患者TNF-α的表达分别是正常对照的1.7、7.0、8.2倍（P＜0.05）;且JAK2V617F突变负荷与TNF-α的表达水平呈线性相关（Pearson r =0.610,R2 =0.372,P=0.005）.结论：TNF-α在BCR-ABL阴性的MPN发病机制中有重要作用,在ET、PV、MF中表达升高且表达量不同,且与JAK2V617F突变呈线性相关.

JAK2V617F Mutation and TNF-α Expression in Myeloproliferative Neoplasms and Their Correlation

This study was aimed to explore the JAK2V617F mutation and TNF-α expression in patients with myeloproliferative neoplasm（ MPN）, and the relation between them so as to provide theoretical basis for clinical practice and target therapy. Sixty-two confirmed BCR-ABL-negative MPN patients and 15 healthy adults were enrolled in this study. The peripheral blood mononuclear cells of the patients and healthy controls were divided into two parts, one part was used to extract DNA, the other one was used to extract mRNA and reverse-transcribe into cDNA. Real-time fluorescent quantitative PCR was used to detect JAK2V617F mutation proportion and the expression level of TNF-α. The results showed that the positive rate of JAK2V617F mutation in MPN patients was 64.52% （40/62）, including 54.28% in essential thrombocythemia （ET） patients （19/35）, 94.74% in polycythemia vera （PV） patients （18/19） and 37.50% in myelofibrosis （MF） （3/8） patients. Mutation proportions of JAK2 V617F in ET, PV and MF patients were 0.838 ±0. 419,4.417 ±0. 658,2. 746 ±2. 009 respectively. The expression of TNF-α in ET, PV and MF patients were higher than that in healthy controls： 1.7,7.0,8.2-fold（ P 〈 0.05 ） respectively. In addition, TNF-α expression was correlated with JAK2 V617F allele burden （ Pearson r = 0.610, R2 = 0. 372, P = 0. 005 ）. It is concluded that TNF-α plays an important role in the pathogenesis of MPN, the TNF-α expression increases and is different in ET, PV and MF patients, which correlates with JAK2 V617F allele burden.