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.
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引用次数: 0
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 in vitro 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.
目的：急性髓系白血病（AML）是髓系干细胞前体的恶性肿瘤。骨髓间充质干细胞衍生的外泌体（BMSC-exos）会影响急性髓性白血病的进展。方法：用从转染了microRNA（miR）-139-5p-mimics、ovβ-catenin或miR-139-5p-抑制剂的骨髓间充质干细胞中提取的外泌体处理体外培养的人AML细胞（MOLM-16、MV-4-11）和正常人造血细胞（GM12878）。显微镜观察 BMSC 形态，油红 O 染色和茜素红 S 染色评估其成脂和成骨分化能力。超速离心法提取 BMSC-外胚层，透射电子显微镜观察其形态。BMSC-exos 通过纳米颗粒追踪分析和 Western 印迹进行鉴定。利用TargetScan数据库预测了miR-139-5p与β-catenin的结合位点，并通过双荧光素酶报告实验进行了验证。RT-qPCR、CCK-8和流式细胞术评估了miR-139-5p和β-catenin的mRNA水平、细胞增殖和凋亡。结果：miR-139-5p 在 AML 细胞系中表达较低，miR-139-5p 过表达会降低 AML 细胞活力/增殖/Bcl-2 水平，提高细胞凋亡/Bax 水平。BMSC-exos 通过 miR-139-5p 抑制 AML 细胞增殖并促进细胞凋亡。随后，上调的β-catenin部分抵消了BMSC-exos中miR-139-5p对AML细胞增殖和凋亡的影响：结论：BMSC-exos通过miR-139-5p抑制β-catenin的表达，限制了AML细胞的增殖并促进了细胞凋亡。
期刊介绍：
Hematology is an international journal publishing original and review articles in the field of general hematology, including oncology, pathology, biology, clinical research and epidemiology. Of the fixed sections, annotations are accepted on any general or scientific field: technical annotations covering current laboratory practice in general hematology, blood transfusion and clinical trials, and current clinical practice reviews the consensus driven areas of care and management.
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