Investigating physiological role of PHF6 protein in T-cell acute lymphoblastic leukemia

Objective: T-cell Acute Lymphoblastic Leukemia is an aggressive type of hematological malignancy that is more common in men than women. It was aimed to elucidate the role of the biological activity of PHF6 in the formation of the disease and the molecular mechanism of the signalling pathway, and has been one of the candidate genes in recent years by playing a tumor suppressor role in T-ALL on immature T cells. We aimed to determine the effect of PHF6 S199 region on protein function in T-ALL and on proliferation, differentiation and migration of T-ALL cells. Method: Following PCR clonning method which performed from human cDNA library into pcDNA3.1A expression vector with PHF6 cDNA sequence, the experiment of changing the S199 region to the A199 and D199 sequences was applied by applying the SDM method to the PHF6 plasmid cloned. PHF6 protein expression vectors transfected into DND-41 cells; Protein expressions evaluated by western blotting, while effects on cell proliferation were determined with MTT wild-type vectors' T-cell differentiation effects evaluated by flow cytometry and cell migration were evaluated by Transwell cell migration method. Results: The ectopic expression of the wild-type PHF6 expression vector transfected in DND-41 cells; expression was increased in the CD4+ direction of the cells, had no effect on the migration of the cells. Reduced Notch1 expression was demonstrated in S199A+ cells. S199A+/D+ mutant vectors have been shown to induced proliferation compared to the PHF6+ cells and control group. Conclusion: This study is the first for working on the cell migration of PHF6 in T-ALL cells in the literature. It was shown that ectopic expression of PHF6; has no effect on cell migration, has the potential to induce T-ALL cells towards CD4+ and S199 site might be significant for Notch1 signalling and cell proliferation.