Nabigol Maryam, Hajipirloo Laya Khodayi, Kuhestani-Dehaghi Bentolhoda, Farsani Mehdi Allahbakhshian
Department of Hematology and Blood Bank, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Hematology and Blood Bank, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Curr Res Transl Med. 2025 Jan-Mar;73(1):103474. doi: 10.1016/j.retram.2024.103474. Epub 2024 Sep 21.
Despite the various therapeutic options introduced for AML treatment, therapy resistance and relapse are still the main obstacles. It is well known that alterations in the bone marrow microenvironment (BMM) play a crucial role in leukemia growth and the treatment failure of AML. Evidence shows that exosomes alter the components of BMM in a way that support leukemia survival, leading to chemoresistance. In this study, we evaluated the effect of AML exosomes on the biological functions of human bone marrow mesenchymal stromal cells (h BM-MSCs), especially alteration in the expression of the JAK/STAT signaling genes, as a leukemia-favoring pathway.
Exosomes were isolated from the HL-60 cell line and characterized using flow cytometry, Transmission Electron Microscopy (TEM), and Dynamic Light Scattering (DLS) technique. The exosome protein content was assessed using a bicinchoninic acid (BCA) protein assay kit in order to determine the concentration of exosomes. Subsequently, MSCs were treated with varying concentrations of AML exosomes, and data was obtained using MTT, cell cycle, apoptosis, and ki67 assays. Additionally, gene expression analysis was conducted through qRT-PCR.
AML exosomes regulated the viability and survival of MSCs in a concentration-dependent manner. The qRT-PCR data revealed that treatment with AML exosomes at a concentration of 50 μg/mL led to a significant upregulation of JAK2, STAT3, and STAT5 genes in MSCs.
Because the JAK/STAT signaling pathway has been shown to play a role in the proliferation and survival of leukemic cells, our results suggest that AML exosomes stimulate MSCs to activate this pathway. This activation may impede AML cell apoptosis, potentially leading to chemoresistance and relapse.
尽管针对急性髓系白血病（AML）治疗引入了多种治疗方案，但治疗耐药和复发仍是主要障碍。众所周知，骨髓微环境（BMM）的改变在白血病生长和AML治疗失败中起关键作用。有证据表明，外泌体以支持白血病存活的方式改变BMM的成分，导致化疗耐药。在本研究中，我们评估了AML外泌体对人骨髓间充质基质细胞（h BM-MSCs）生物学功能的影响，特别是作为白血病有利途径的JAK/STAT信号基因表达的改变。
从HL-60细胞系中分离外泌体，并使用流式细胞术、透射电子显微镜（TEM）和动态光散射（DLS）技术进行表征。使用二辛可宁酸（BCA）蛋白质测定试剂盒评估外泌体蛋白质含量，以确定外泌体的浓度。随后，用不同浓度的AML外泌体处理间充质干细胞，并使用MTT、细胞周期、凋亡和ki67测定获得数据。此外，通过qRT-PCR进行基因表达分析。
AML外泌体以浓度依赖的方式调节间充质干细胞的活力和存活。qRT-PCR数据显示，用50μg/mL浓度的AML外泌体处理导致间充质干细胞中JAK2、STAT3和STAT5基因显著上调。
由于JAK/STAT信号通路已被证明在白血病细胞的增殖和存活中起作用，我们的结果表明AML外泌体刺激间充质干细胞激活该通路。这种激活可能会阻碍AML细胞凋亡，潜在地导致化疗耐药和复发。
