Rassaei Neda, Abbaszade Dibavar Mahnoosh, Soleimani Masoud, Atashi Amir, Mohammadi Mohammad Hossein, Allahbakhshian Farsani Mehdi, Shahsavan Shaghayegh
Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University,Tehran, Iran.
Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Iran J Basic Med Sci. 2023 Mar;26(3):295-300. doi: 10.22038/IJBMS.2023.66903.14675.
Microvesicles (MVs) are small membrane-bound particles that act as a vehicle to transfer their contents, such as proteins, RNAs, and miRNAs, to the target cells, making them undergo several changes. Depending on the origin and the target cell, MVs may cause cell survival or apoptosis. This study investigated the effects of MVs released from the leukemic K562 cell line on the human bone marrow mesenchymal stem cells (hBM-MSCs) to evaluate changes in the survival or apoptosis of the cells in an system.
In this experimental study, we added the isolated MVs from the K562 cell line to hBM-MSCs, and after three and then seven days, subsequently cell count, cell viability, transmission electron microscopy, tracing MVs by carboxyfluorescein diacetate, succinimidyl ester (CFSE) solution, flow cytometry analysis for Annexin-V/PI staining and qPCR for the evaluation of 2, , and expression were carried out. On the 10 day of the culture, hBM-MSCs were examined by Oil red O and Alizarin Red staining to evaluate their differentiation into adipocytes and osteoblasts.
There was a significant decrease in cell viability and and expression; however, was significantly upregulated in the hBM-MSCs compared to control groups. Annexin-V/PI staining results also showed the apoptotic effects of K562-MVs on hBM-MSCs. Moreover, the differentiation of hBM-MSCs into adipocytes and osteoblasts was not observed.
MVs from the leukemic cell line could affect the viability of normal hBM-MSCs and induce cell apoptosis.
微泡(MVs)是一种小的膜结合颗粒,可作为载体将其内容物(如蛋白质、RNA和微小RNA)转移至靶细胞,使其发生多种变化。根据其来源和靶细胞的不同,微泡可能导致细胞存活或凋亡。本研究调查了白血病K562细胞系释放的微泡对人骨髓间充质干细胞(hBM-MSCs)的影响,以评估在一个系统中细胞存活或凋亡的变化。
在本实验研究中,我们将从K562细胞系分离出的微泡添加到hBM-MSCs中,在三天后和七天后,分别进行细胞计数、细胞活力检测、透射电子显微镜观察、用羧基荧光素二乙酸琥珀酰亚胺酯(CFSE)溶液追踪微泡、进行膜联蛋白V/碘化丙啶(Annexin-V/PI)染色的流式细胞术分析以及用于评估2、、和表达的定量聚合酶链反应(qPCR)。在培养的第10天,通过油红O和茜素红染色检测hBM-MSCs,以评估其向脂肪细胞和成骨细胞的分化情况。
细胞活力以及和表达显著降低;然而,与对照组相比,在hBM-MSCs中显著上调。Annexin-V/PI染色结果也显示K562-MVs对hBM-MSCs具有凋亡作用。此外,未观察到hBM-MSCs向脂肪细胞和成骨细胞的分化。
白血病细胞系来源的微泡可影响正常hBM-MSCs的活力并诱导细胞凋亡。