Subotički Tijana, Mitrović Ajtić Olivera, Beleslin-Čokić Bojana B, Nienhold Ronny, Diklić Miloš, Djikić Dragoslava, Leković Danijela, Bulat Tanja, Marković Dragana, Gotić Mirjana, Noguchi Constance T, Schechter Alan N, Skoda Radek C, Čokić Vladan P
Institute for Medical Research, University of Belgrade, Belgrade, Serbia.
Clinic for Endocrinology, Diabetes and Metabolic Diseases, Genetic Laboratory, Clinical Center of Serbia, Belgrade, Serbia.
Mol Carcinog. 2017 Feb;56(2):567-579. doi: 10.1002/mc.22517. Epub 2016 Jul 8.
It has been shown that angiogenesis and inflammation play an important role in development of most hematological malignancies including the myeloproliferative neoplasm (MPN). The aim of this study was to investigate and correlate the levels of key angiogenic molecules such as hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) in peripheral blood and bone marrow cells of MPN patients, along with JAK2V617F mutation allele burden and effects of therapy. HIF-1α and VEGF gene expression were decreased, while eNOS mRNA levels were increased in granulocytes of MPN patients. Furthermore, positively correlated and increased VEGF and eNOS protein levels were in negative correlation with HIF-1α levels in granulocytes of MPN patients. According to immunoblotting, the generally augmented angiogenic factors demonstrated JAK2V617F allele burden dependence only in granulocytes of PMF. The angiogenic factors were largely reduced after hydroxyurea therapy in granulocytes of MPN patients. Levels of eNOS protein expression were stimulated by Calreticulin mutations in granulocytes of essential thrombocythemia. Immunocytochemical analyses of CD34 cells showed a more pronounced enhancement of angiogenic factors than in granulocytes. Increased gene expression linked to the proinflammatory TGFβ and MAPK signaling pathways were detected in CD34 cells of MPN patients. In conclusion, the angiogenesis is increased in several cell types of MPN patients supported by the transcriptional activation of inflammation-related target genes, and is not limited to bone marrow stroma cells. It also appears that some of the benefit of hydroxyurea therapy of the MPN is mediated by effects on angiogenic factors. © 2016 Wiley Periodicals, Inc.
研究表明，血管生成和炎症在包括骨髓增殖性肿瘤（MPN）在内的大多数血液系统恶性肿瘤的发展中起着重要作用。本研究的目的是调查MPN患者外周血和骨髓细胞中关键血管生成分子如缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）和内皮型一氧化氮合酶（eNOS）的水平，并将其与JAK2V617F突变等位基因负荷及治疗效果相关联。MPN患者粒细胞中HIF-1α和VEGF基因表达降低，而eNOS mRNA水平升高。此外，MPN患者粒细胞中VEGF和eNOS蛋白水平呈正相关且升高，与HIF-1α水平呈负相关。根据免疫印迹法，一般增强的血管生成因子仅在原发性骨髓纤维化（PMF）患者的粒细胞中表现出对JAK2V617F等位基因负荷的依赖性。MPN患者粒细胞经羟基脲治疗后，血管生成因子大幅减少。在原发性血小板增多症患者的粒细胞中，钙网蛋白突变刺激了eNOS蛋白表达水平。对CD34细胞的免疫细胞化学分析显示，血管生成因子的增强比粒细胞中更明显。在MPN患者的CD34细胞中检测到与促炎TGFβ和MAPK信号通路相关的基因表达增加。总之，MPN患者的几种细胞类型中血管生成增加，这是由炎症相关靶基因的转录激活所支持的，且不限于骨髓基质细胞。似乎MPN患者接受羟基脲治疗的一些益处是由对血管生成因子的作用介导的。© 2016威利期刊公司
