FOXO3 Expression as a Predictive Factor in Developing Chronic Myeloid Leukemia Imatinib Resistance

Background and Aim: Mesenchymal stem cells have a long-term clinicalapplication and widely used in autoimmune disease and regenerativemedicine. However, some MSCs derived cytokines such as TGF-β couldhave a dual role in suppression or progression of the disease. Fibroblastactivation and extracellular matrix production are two key features ofwound healing and contrivers in fibrogenesis. In this study we considerthe role of MSC derived condition media as a source of MSc derivedsecretome on human fibroblast activation.Methods: To isolate human stem cells, bone marrow was collected froma healthy donor. After separating cells and cultured on DMEM/ FBS10%,cells with fibroblast shape were isolated as mesenchymal stem cells.To characterize the MSC, flow cytometric assays for surface receptormolecule expression and differentiation were performed. Human foreskinfibroblast (HFF) was cultured under standard condition. To collect MSCCM,cells were cultured in the serum-free media for 72 hours. HFF cellswere plated at a seeding density of 2×105 cells/well in a 6-well plateand after 24 hours they were treated with MSC-CM and exogenous TGFbeta was applied as a positive control. Total RNA from samples wasisolated and purified using the RNeasy mini kit (Qiagen). Collagen andalpha-SMA expression as a fibrotic marker was determined by RT-PCR.Expression levels of all genes were analyzed according to comparative analysis method by Rest and GraphPad software.Results: Flow cytometry and differentiation methods identified cellsas MSc with purity of 98%. MSc markers including CD73, CD90, andCD105 were positive and CD14, CD34 and CD 45 reported as negative.After 7 2h treatment of HFF by MSc conditioned media, alpha-SMAshowed 2-fold upregulation in comparison with control. Collagen III asa second fibrotic marker also showed a significant increase of mRNA(P<0.05). TGF-β also showed similar results compared to TGF-B andcondition media, it was shown; MSC-CM unregulated the expression ofcollagen III and alpha-SMA target genes which are involved in fibrosisand known as a myofibroblast marker.Conclusion: MSCs assumed to be a reliable source of stem cells inclinical application. Our data demonstrate that MSC produces cytokinesthat stimulate matrix production. It will be essential to determine whetherthese factors can play a role in attempts to use MSC for therapeuticapproaches. These data suggest that MSC can significantly stimulate thedifferentiation of fibroblast cells to myofibroblasts which has a pivotalrole in the onset of a Fibrogenesis. However, there is a deal to be learnedabout the mechanism through which MSC play a role in the developmentof the disease.

کلیدواژه ها:

MSC-secretome ، Fibroblast ، TGF-B ، αSMA ، Collagen III ، Myofibroblast