IFN-α2b对JAK2V617F突变的骨髓增殖性肿瘤COX-2表达及血管新生的影响

Effect of IFN-α2b on COX-2 and Angiogenesis in JAK2V617F Mutation Myeloproliferative Neoplasms

目的研究干扰素-alpha-2b(IFN-α2b)对骨髓增殖性肿瘤(MPN)患者JAK2激酶、环氧化酶-2(COX-2)及微血管密度的影响及人红白血病HEL细胞系中JAK2V617F与COX-2之间的关系。方法收集在保定市第一医院接受初次治疗的42例有JAK2V617F突变的MPN患者,其中17例患者为IFN-α2b治疗组(给予IFN-α2b肌肉注射治疗),25例为初治组(未行治疗)。另取同期10例特发性免疫性血小板减少性紫癜(ITP)患者作为对照。采用实时荧光定量聚合酶链反应(qRT-PCR)检测MPN患者JAK2V617F/JAK2突变量,免疫组化检测MPN患者和ITP患者骨髓病理组织p-JAK2、COX-2及CD105标记的微血管密度(MVD)。用不同浓度IFN-α2b作用于HEL细胞系,CCK-8检测细胞增殖抑制率,流式细胞术检测凋亡率,Transwell小室检测细胞迁移能力,半定量PCR检测HEL细胞中JAK2、COX-2mRNA表达水平,Western blot检测p-JAK2、COX-2蛋白表达水平。结果初治组患者p-JAK2、COX-2表达水平及MVD高于对照组,IFN-α2b治疗后患者p-JAK2、COX-2及MVD表达水平减低。不同剂量IFN-α2b作用HEL细胞48h,细胞增殖抑制率和细胞凋亡率随其剂量增加逐渐上升,JAK2及COX-2mRNA及p-JAK2、COX-2蛋白表达随其剂量增加逐渐降低。细胞迁移实验结果发现加入0.5×104 U/L IFN-α2b处理HEL细胞24h,迁移至下室的细胞数低于对照组(P<0.05)。结论 IFN-α2b通过调控JAK2信号通路抑制COX-2及MPN患者血管新生。

Objective To investigate the influence of interferon-alpha-2b（IFN-α2b）with JAK2 kinase,COX-2and microvessel density in patients of MPN and the relation of JAK2V617 Fand COX-2in human erythroleukemia cell line（HEL）cells.Methods Forty-two cases of MPN patients with JAK2V617 F mutation of initial treatment were collected from the Frist hospital of Baoding,including the IFN-α2btreatment group with 17 cases and untreated group with 25 cases.10cases of idiopathic immune thrombocytopenic purpura（ITP）patients synchronization were enrolled as controls.JAK2V617F/JAK2 mutation burden of MPN patients was detected by real time PCR（qRTPCR）;the expression levels of p-JAK2,COX-2 and microvascular density（MVD） marked with CD105 inpathological tissues of bone marrow in patients of MPN and ITP were detected by immunohistochemistry.The HEL cells were treated with different concentrations of IFN-α2b.The cell proliferation inhibition rate was calculated by CCK-8test;the apoptosis rate was detected by flow cytometry;cell migration ability was tested by transwell chambers.JAK2 and COX-2mRNA were detected by semi-quantitative PCR;p-JAK2 and COX-2protein in HEL cells were detected by Western blotting.Results The expression levels of p-JAK2,COX-2protein and MVD in untreated group were significantly higher than those of control groups.p-JAK2,COX-2and MVD levels were significantly reduced in patients treated with IFN-α2b.Cell growth inhibition rates and apoptosis rates raise up by dose of IFN-α2bin HEL cells at 48 h.The mRNA expression levels of JAK2 and COX-2as well as protein expression levels of p-JAK2 and COX-2had a decreasing tendency with the increase of IFN-α2bconcentration at48 h.The migration capacity level of HEL cells which treated with 0.5×104 U/L IFN-α2bafter 24 hwas lower than that of control group.Conclusion Angiogenesis of MPN and COX-2 were inhibited by IFN-α2bwhich regulates JAK2 signal pathway.