The Study Of Expression And Co-relation Between The JAK2V617F And IL-6, TNF-α, IL-32, IL-37 In Myeloproliferative Neoplasms

Objective:This study was aimed to explore the JAK2V617F mutation and TNF-a, IL-6, IL-32, IL-37 expression in patients with myeloproliferative neoplasms (MPN), and investigate the relation between them so as to provide theoretical basis for clinical practice and prove the theory that chronic inflammation is considered of major importance in the development of MPN.Methods:According to the latest revision of WHO diagnostic criteria, seventy-nine confirmed BCR-ABL-negative MPN patients and fifteen healthy adults were enrolled in this study.5ml peripheral blood of every patient were collected and abstracted the mononuclear cells. The peripheral blood mononuclear cells of the patients and healthy controls were divided into two parts, one part was extracted DNA, the other one was extracted mRNA and reverse-transcribed into cDNA. Real-time fluorescent quantitative PCR was used to detect JAK2V617F allele burden and the expression level of TNF-a, IL-6, IL-32, IL-37. Correlations between the JAK2V617F allele burden and the expression level of TNF-a, IL-6, IL-32, IL-37 were analysed by Statistical Packages for the Social Sciences v21.0 software, P value<0.05 was considered as statistically significant.Results:The positive rate of JAK2V617F mutation in MPN patients was 63.29%(50/79), including 51.11% in essential thrombocythemia (ET) patients (23/45),95.83% in polycythemia vera (PV) patients (23/24) and 40.00% in myelofibrosis (MF) patients (4/10). Allele burden of JAK2V617F in ET, PV and MF patients were 22.55%±4.59%, 67.90%±7.29%,55.38%±6.55% respectively. The gene expression of TNF-a, IL-6, IL-32, IL-37 in ET, PV and MF were higher than healthy controls,1.81,6.95, 8.03-fold(P<0.05),2.92,5.50,35.05-fold(P<0.05),0.28,0.41,0.68-fold(P<0.05), 14.29,19.75,6.90-fold(P<0.05), respectively. In addition, the JAK2V617F allele burden was correlated with TNF-a expression(Pearson r=0.61, R2=0.37, P<0.01) and IL-6 expression(Pearson r=0.45, R2=0.21, P<0.01) respectively, but not with IL-32 and IL-37, besides, the correlation between TNF-a and IL-6 is significant(Pearson r=0.60, R2=0.36,P<0.01).Conclusion:1. JAK2V617F allele burden was high in BCR-ABL negative MPN patients, so it can be used as an important indicator of MPN diagnosis. The allele burden of JAK2V617F was significantly lower for ET than for PV and MF, which depicted a biologic continuum from early disease to the advanced myelofibrosis stage.2. MPN patients over-expressed TNF-a, IL-6, IL-37, but the IL-32 expression was markedly reduced. Therefore, inflammation-linked cytokines are required for the development of MPN.3. JAK2V617F mutation load was correlated with the mRNA expression of TNF-a and IL-6, but not with IL-32 and IL-37, indicating that TNF-a and IL-6 may affect JAK2V617F mutation rate via JAK2V617F signal transduction pathway, which provide new thoughts for diagnosis and treatment of MPN. Apart from this, TNF-a had a linear correlation with IL-6 expression.