Objective: Patients with myeloproliferative neoplasms (MPN) are at an increased risk of developing thrombosis which leads to significant levels of morbidity and mortality. Although this propensity for thrombosis has been attributed to increased blood counts, other factors such as endothelial cell (EC) dysfunction likely play an important role. Previous studies have provided evidence of EC involvement by JAK2V617F which is known to be present in blood cells of 50-95% of patients with MPN. Furthermore, the morphological and functional changes of this mutation, a single nucleotide mutation in the cytosolic tyrosine kinase, Janus kinase 2 (JAK2) has not been shown in EC. In addition, the role of epigenetic factors including suppressors of cytokine signaling (SOCS) proteins is also unknown for MPN. In this study, the effects of the JAK2V617F mutation on EC and the SOCS 1-4 genes expression and their methylation profiles were investigated. Materials and Methods: The in vitro transduction method of green fluorescent protein (GFP) labeled lentivirus carrying the JAK2 (wild form) or JAK2V617F mutation into human cord vein endothelial cell (HUVEC) was applied. GFP was determined and analyzed with the flow cytometer. Afterwards, the genetically modified EC population was isolated, gating for GFP signal by cell sorter and then DNA and RNA isolations were performed. Gene expressional changes of SOCS1-4 genes were determined in real time RT-PCR and methylation specific PCR was performed with bisulfite modification of DNA. Results: The successful infection of JAK2, JAK2V617F and GFP vectors into EC were achieved. The comparison of the genetically modified EC revealed no morphological differences. A significant increase in the expressions of SOCS1-4 were detected in mutant EH compared to JAK2 and GFP. It was determined that methylation had no effect on suppression of SOCS1-4 expression detected in GFP and JAK2 EC. Conclusion: Future studies will further enlighten the effects of JAK2V617F on EC and its role in thrombotic events.
|Destekleyen Kurum||İstanbul Üniversitesi Bilimsel Araştırma Projeleri Birimi (BAP) tarafından desteklenmiştir.|
|Proje Numarası||Proje No: 27570|
: 6 Eylül 2020
|APA||Göksu, A , Heki̇moğlu, H , Sözer Tokdemi̇r, S . (2020). Miyeloproliferatif Neoplazilerde JAK2V617F Mutasyonunun Endotel Hücresine Etkisi ve SOCS1-4 Gen Anlatımlarına Yansıması . Sağlık Bilimlerinde İleri Araştırmalar Dergisi , 3 (3) , 135-147 . Retrieved from https://dergipark.org.tr/tr/pub/sabiad/issue/57700/790117|