骨髓间充质干细胞来源的外泌体介导 microRNA-139-5p 调控 β-连环蛋白在调节急性髓系白血病细胞增殖和凋亡中的作用机制。

Mechanism of bone-marrow mesenchymal stem cell-derived exosomes mediating microRNA-139-5p to regulate β-catenin in the modulation of proliferation and apoptosis of acute myeloid leukemia cells.

机构信息

Division of Hematology & Oncology, Department of Geriatrics, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, People's Republic of China.

出版信息

Hematology. 2024 Dec;29(1):2428482. doi: 10.1080/16078454.2024.2428482. Epub 2024 Nov 21.

DOI:10.1080/16078454.2024.2428482

Abstract

OBJECTIVE

Acute myeloid leukemia (AML) stands out as a malignancy of the stem cell precursors of the myeloid lineage. Bone-marrow mesenchymal stem cell-derived exosomes (BMSC-exos) affect AML progression. We explored the effects and mechanism of BMSC-exos on AML cell proliferation and apoptosis.

METHODS

Human AML cells (MOLM-16, MV-4-11) and normal human hematopoietic cells (GM12878) cultured were treated with exos extracted from BMSCs that transfected with microRNA (miR)-139-5p-mimics, ovβ-catenin, or miR-139-5p-inhibitor. BMSC morphology was observed by a microscopy, and its adipogenic and osteogenic differentiation abilities were assessed by oil red O staining and alizarin red S staining. BMSC-exos were extracted by ultracentrifugation, and their morphology was observed by a transmission electron microscopy. BMSC-exos were identified by nanoparticle tracking analysis and Western blot. The binding sites between miR-139-5p and β-catenin were predicted by TargetScan database, and then validated by dual-luciferase reporter assay. mRNA levels of miR-139-5p and β-catenin, cell proliferation, and apoptosis were evaluated by RT-qPCR, CCK-8, and flow cytometry. The expressions of CD63, CD81, TSG101, and GRP94 and the proteins of β-catenin, Bax, and Bcl-2 were determined by Western blot.

RESULTS

miR-139-5p was poorly expressed in AML cell lines. miR-139-5p overexpression reduced AML cell viability/proliferation/Bcl-2 level, and raised apoptosis/Bax level. BMSC-exos repressed AML cell proliferation and promoted apoptosis via miR-139-5p. miR-139-5p targeted to inhibit β-catenin expression. Subsequently, up-regulated β-catenin partially counteracted the effects of miR-139-5p in BMSC-exos on AML cell proliferation and apoptosis.

CONCLUSION

BMSC-exos targeted to repress β-catenin expression by miR-139-5p, limited AML cell proliferation and facilitated apoptosis.

摘要

目的

急性髓系白血病（AML）是骨髓髓系前体细胞的恶性肿瘤。骨髓间充质干细胞衍生的外泌体（BMSC-exos）影响 AML 的进展。我们探讨了 BMSC-exos 对 AML 细胞增殖和凋亡的影响及机制。

方法

用转染 microRNA（miR）-139-5p 模拟物、ovβ-catenin 或 miR-139-5p 抑制剂的 BMSC 提取的外泌体处理人 AML 细胞（MOLM-16、MV-4-11）和正常人造血细胞（GM12878）。通过显微镜观察 BMSC 形态，通过油红 O 染色和茜素红 S 染色评估其成脂和成骨分化能力。通过超速离心提取 BMSC-exos，通过透射电子显微镜观察其形态。通过纳米颗粒跟踪分析和 Western blot 鉴定 BMSC-exos。通过 TargetScan 数据库预测 miR-139-5p 和β-catenin 的结合位点，然后通过双荧光素酶报告基因检测验证。通过 RT-qPCR、CCK-8 和流式细胞术评估 miR-139-5p 和β-catenin 的 mRNA 水平、细胞增殖和凋亡。通过 Western blot 测定 CD63、CD81、TSG101 和 GRP94 的表达以及β-catenin、Bax 和 Bcl-2 的蛋白表达。

结果

miR-139-5p 在 AML 细胞系中表达水平较低。miR-139-5p 过表达降低 AML 细胞活力/增殖/Bcl-2 水平，增加凋亡/Bax 水平。BMSC-exos 通过 miR-139-5p 抑制 AML 细胞增殖并促进凋亡。miR-139-5p 靶向抑制β-catenin 表达。随后，上调的β-catenin 部分抵消了 BMSC-exos 中 miR-139-5p 对 AML 细胞增殖和凋亡的作用。