The effect of extracellular genome materials on hematopoietic stem cells

Aim:Main aim of thesis is successfully transfected HSC(Hematopoietic stem cells) and determine cellular function changes. Extracellular genome materials(EGM) found in all biological fluids which can released from cells and integrated into neighboring and distal cells.EGMs may carry oncogenes, they can oncogenically transform normal cells.Because of roles in the interaction between cells, EGMs were used for transfection in this study and it was show that HSCs can differentiate into cancer stem cells. Methods:Cord blood was collected from donors and mononuclear cells were isolated. Then, CD34+/- cells were sorted. HEL cell was used because it carries the JAK2-V617F biallelic and the K562 cell was BCR/ABL1 positive. CD34+/- cells were transfected with serum collected from these cultures. In order to detect changes in transfected cells, ımmunophenotyping was performed with progenitor cell colony(PCC), apoptosis and proliferation assays. Results: HEL serum transfected CD34+/- cells were detected the JAK2-V617F mutation monoallelic in 24h.When the incubate period was 48 hours, HSCs were JAK2-V617 mutation biallelic. After transfection, all samples(100%) of K562 serum transfected HSCs were detected BCR/ABL1 positive. While JAK2-V617F mutation was shown at a rate of 50% in transfected PCC; BCR/ABL1 was shown at a rate of 6%. after transfection, the cell function analysis was performed. After transfection, the cell function analysis was increased as apoptosis of CD34- cell, pSTAT5 in HEL/CD34+/- , lympocyte count in K562 serum/CD34+/-. Conclusion:Our findings support literature that EGMs can transfer the oncogenes, carry from the cells they are released to different cells laterally and change biological properties of cell by giving these cells oncogenic properties. In this study, we have shown for the first time that it is possible to integrate the JAK2-V617F mutation and BCR/ABL1 fusion genes into HSCs via EGMs. These data show that hdGMs play a role in intercellular communication and contribute to genometastasis.