Student Research Committee, Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Pediatric Congenital Hematologic Disorders Research Center, Research Institute for Children's Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Mol Biol Rep. 2024 Jun 14;51(1):749. doi: 10.1007/s11033-024-09674-4.
Background The incidence of various types of cancers, including leukemia, is on the rise and many challenges in both drug resistance and complications related to chemotherapy appeared. Recently, the development and application of extracellular vesicles (EV) such as exosomes in the management of cancers, especially leukemia, holds great significance. In this article, we extracted exosomes from NALM6 cells and assessed their regulatory effects on proliferation and apoptosis in mesenchymal stem cells (MSCs). Method and result We first verified the exosomes using various techniques, including flow cytometry, transient electron microscopy, dynamic light scattering (DLS), and BCA protein assay. Then MTT analysis and flowcytometry (apoptosis and cell cycle assay) besides gene expressions were employed to determine the state of MSC proliferations. The results indicated that exosome-specific pan markers like CD9, CD63, and CD81 were present. Through DLS, we found out that the mean size of the exosomes was 89.68 nm. The protein content was determined to be 956.292 µg/ml. Analysis of MTT, flow cytometry (cell cycle and apoptosis assay), and RT-qPCR showed that in the dose of 50 µg/ml the proliferation of MSCs was increased significantly (p-value < 0.05). Conclusion All these data showed that exosomes use several signaling pathways to increase the MSCs' proliferation and drug resistance, ultimately leading to high mortalities and morbidities of acute lymphoblastic leukemia.
包括白血病在内的各种癌症的发病率呈上升趋势,在药物耐药性和与化疗相关的并发症方面出现了许多挑战。最近,细胞外囊泡(EV)如外泌体在癌症,尤其是白血病的治疗中的开发和应用具有重要意义。在本文中,我们从 NALM6 细胞中提取外泌体,并评估其对间充质干细胞(MSCs)增殖和凋亡的调节作用。
我们首先使用各种技术,包括流式细胞术、瞬态电子显微镜、动态光散射(DLS)和 BCA 蛋白测定法,验证了外泌体。然后,我们使用 MTT 分析和流式细胞术(凋亡和细胞周期分析)以及基因表达来确定 MSC 增殖状态。结果表明,存在外泌体特异性泛标志物,如 CD9、CD63 和 CD81。通过 DLS,我们发现外泌体的平均大小为 89.68nm。蛋白质含量确定为 956.292µg/ml。MTT 分析、流式细胞术(细胞周期和凋亡分析)和 RT-qPCR 分析表明,在 50µg/ml 的剂量下,MSCs 的增殖显著增加(p 值<0.05)。
所有这些数据表明,外泌体通过几种信号通路增加 MSCs 的增殖和耐药性,最终导致急性淋巴细胞白血病的高死亡率和发病率。