Jalilivand Sahar, Nabigol Maryam, Bakhtiyaridovvombaygi Mehdi, Gharehbaghian Ahmad
Department of Laboratory Hematology and Blood Bank, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Student Research Committee, Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Blood Res. 2024 Dec 20;59(1):43. doi: 10.1007/s44313-024-00051-5.
Despite advances in the treatment of acute myeloid leukemia (AML), refractory forms of this malignancy and relapse remain common. Therefore, development of novel, synergistic targeted therapies are needed urgently. Recently, mesenchymal stem cells (MSCs) have been shown to be effective in treating various diseases, with most of their therapeutic outcomes attributed to their exosomes. In the current study, we investigated the effects of bone marrow mesenchymal stem cell (BM-MSC) exosomes on the expression of the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling genes involved in AML pathogenesis.
Exosomes were isolated from BM-MSCs and confirmed using transmission electron microscopy, dynamic light scattering, and flow cytometry. Subsequently, the exosome concentration was estimated using the bicinchoninic acid assay, and HL-60 cells were cocultured with 100 µg/mL of BM-MSC exosomes. Finally, the JAK2, STAT3, and STAT5 expression levels were analyzed using qRT-PCR.
The exosome characterization results confirmed that most isolated nanoparticles exhibited a round morphology, expressed CD9, CD63, and CD81, which are specific protein markers for exosome identification, and ranged between 80 and 100 nm in diameter. Furthermore, qRT-PCR analysis revealed a significant downregulation of JAK2, STAT3, and STAT5 in HL-60 cells treated with 100 μg/mL of BM-MSC exosomes.
Since JAK/STAT signaling contributes to AML survival, our findings suggest that the downregulation of JAK/STAT genes by BM-MSC exosomes in leukemic cells may aid in designing a potent therapeutic strategy for AML treatment.
尽管急性髓系白血病（AML）的治疗取得了进展，但这种恶性肿瘤的难治性形式和复发仍然很常见。因此，迫切需要开发新型的、具有协同作用的靶向治疗方法。最近，间充质干细胞（MSCs）已被证明在治疗各种疾病方面有效，其大部分治疗效果归因于其外泌体。在本研究中，我们调查了骨髓间充质干细胞（BM-MSC）外泌体对参与AML发病机制的Janus激酶/信号转导和转录激活因子（JAK/STAT）信号基因表达的影响。
从BM-MSCs中分离出外泌体，并使用透射电子显微镜、动态光散射和流式细胞术进行确认。随后，使用二辛可宁酸测定法估计外泌体浓度，并将HL-60细胞与100μg/mL的BM-MSC外泌体共培养。最后，使用qRT-PCR分析JAK2、STAT3和STAT5的表达水平。
外泌体表征结果证实，大多数分离的纳米颗粒呈圆形形态，表达CD9、CD63和CD81，这些是用于外泌体鉴定的特异性蛋白质标志物，直径在80至100nm之间。此外，qRT-PCR分析显示，用100μg/mL的BM-MSC外泌体处理的HL-60细胞中JAK2、STAT3和STAT5显著下调。
由于JAK/STAT信号传导有助于AML的存活，我们的研究结果表明，BM-MSC外泌体在白血病细胞中对JAK/STAT基因的下调可能有助于设计一种有效的AML治疗策略。
