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骨髓间充质干细胞分泌的细胞因子可促进 K562 细胞系的细胞凋亡和改变细胞周期分布,可作为细胞移植中的临床药物。

Cytokines secreted from bone marrow derived mesenchymal stem cells promote apoptosis and change cell cycle distribution of K562 cell line as clinical agent in cell transplantation.

机构信息

Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

PLoS One. 2019 Apr 22;14(4):e0215678. doi: 10.1371/journal.pone.0215678. eCollection 2019.

Abstract

Mesenchymal stem cells (MSCs) are of special interest due their potential clinical use in cell-based therapy. Therapies engaging MSCs are showing increasing promise in the cancer treatment and anticancer drug screening applications. A multitude of growth factors and cytokines secreted from these cells are known to give such multifunctional properties, but details of their role are yet to be absolutely demonstrated. In this study, we have evaluated the influence of BMSCs on K562 cell line as chronic myeloid leukemia (CML) cells, with the use of a cytokine antibody array recognizing 34 cytokines. For this purpose, BMSCs were isolated and co-cultured with K562 cells; thereafter, cultured K562 alone and co-cultured K562 with BMSCs (10:1) were collected at day 7 and subjected to cell cycle distribution assay as well as annexin/PI analysis and Ki/caspase-3 assay for apoptosis assessment. In the following, the gene and protein expression levels of BAX and BCL-2 as pro- and anti-apoptotic agents were investigated. Furthermore, after 7 days' treatment, culture medium was collected from both control and experimental groups for cytokine antibody array. It was found that BMSCs resulted in a robust increase in the number of cells at G0/G1 phase and arrest the G0/G1 phase as well as significantly inducing late apoptosis in K562 cells. The significant presence of TIMP-1 (tissue inhibitor of metalloproteinases-1), and moderate elevated signals for CINC-1 (cytokine-induced neutrophil chemoattractant-1) were obvious in the co-cultured conditioned media, but no significant increase was found in 32 other cytokines. It is concluded that co-culture of BMSCs with K562 cells could secrete a substantial amount of TIMP-1 and CINC-1. These cytokines could be involved in the inhibition of the K562 cell proliferation via BAX and caspase-3 cascade pathways.

摘要

间充质干细胞(MSCs)因其在细胞治疗中的潜在临床应用而备受关注。涉及 MSCs 的疗法在癌症治疗和抗癌药物筛选应用中显示出越来越大的前景。这些细胞分泌的多种生长因子和细胞因子被认为赋予了它们多功能特性,但它们的作用细节尚未得到绝对证明。在这项研究中,我们评估了 BMSCs 对慢性髓系白血病(CML)细胞系 K562 细胞的影响,使用了一种识别 34 种细胞因子的细胞因子抗体阵列。为此,我们分离了 BMSCs 并与 K562 细胞共培养;此后,在第 7 天收集单独培养的 K562 细胞和共培养的 K562 与 BMSCs(10:1)细胞,并进行细胞周期分布分析以及 Annexin/PI 分析和 Ki/caspase-3 分析以评估细胞凋亡。接下来,我们研究了促凋亡和抗凋亡剂 BAX 和 BCL-2 的基因和蛋白表达水平。此外,在 7 天的治疗后,从对照组和实验组收集培养基进行细胞因子抗体阵列分析。结果发现,BMSCs 导致 G0/G1 期细胞数量显著增加,并使 G0/G1 期停滞,同时显著诱导 K562 细胞晚期凋亡。在共培养的条件培养基中,TIMP-1(金属蛋白酶组织抑制剂-1)的存在明显,CINC-1(细胞因子诱导的中性粒细胞趋化因子-1)的信号也适度升高,但其他 32 种细胞因子的信号没有明显增加。因此,BMSCs 与 K562 细胞共培养可分泌大量 TIMP-1 和 CINC-1。这些细胞因子可能通过 BAX 和 caspase-3 级联途径参与抑制 K562 细胞增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8019/6476492/2f3094f11be9/pone.0215678.g001.jpg

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