Introduction: Acute myeloid leukemia (AML) is a predominant blood malignancy with high mortality and severe morbidity. AML is affected by microRNAs (miRNAs) loaded in exosomes derived from bone marrow mesenchymal stem cells (BM-MSCs). MiR-425-5p has been reported to participate in different cancer models. However, the function of BM-MSCs-derived exosomal miR-425-5p in AML is unclear.
Methods: The expression of miR-425-5p was measured by qRT-PCR in clinical AML samples. The immunophenotype of BM-MSCs was analyzed using antibodies against CD44, CD90, and CD105. The exosome was isolated from BM-MSCs. The effect of BM-MSCs-derived exosomal miR-425-5p on AML was analyzed by CCK-8 assay, Edu assay, transwell assay, flow cytometry in AML cells. qRT-PCR, luciferase reporter gene assay and Western blot analysis were also conducted in AML cells.
Results: The expression levels of miR-425-5p were decreased in CD34 + CD38-AML cells from primary AML patients compared to that from the bone marrow of healthy cases, and were reduced in exosomes from AML patients compared that from healthy cases. Similarly, miR-425-5p was also down-regulated in AML cell lines compared with BM-MSCs. MiR-425-5p was able to express in exosomes from BM-MSCs. CCK-8, Edu, transwell assay and flow cytometry analysis revealed that BM-MSCs-derived exosomal miR-425-5p significantly inhibited cell viability, Edu positive cells, invasion and migration, and induced apoptosis of AML cells. Meanwhile, the expression levels of cleaved PARP and cleaved caspase3 were increased by BM-MSCs-derived exosomal miR-425-5p in cells. MiR-425-5p inhibited the expression of Wilms tumor 1-associated protein (WTAP). Moreover, overexpression of WTAP could reverse the miR-425-5p-induced inhibition effect on AML cell proliferation, apoptosis, migration and invasion.
Conclusion: BM-MSCs-derived exosomal miR-425-5p inhibits proliferation, invasion and migration of AML cells and induced apoptosis of AML cells by targeting WTAP. Therapeutically, BM-MSCs-derived exosomal miR-425-5p may serve as a potential target for AML therapy.

Keywords: AML, BM-MSCs, exosome, miR-425-5p, WTAP


中文翻译：

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骨髓间充质干细胞衍生的外泌体 miR-425-5p 通过靶向 WTAP 抑制急性髓系白血病细胞增殖、凋亡、侵袭和迁移

简介：急性髓性白血病（AML）是一种主要的血液恶性肿瘤，具有高死亡率和严重的发病率。AML 受到来自骨髓间充质干细胞 (BM-MSC) 的外泌体中加载的 microRNA (miRNA) 的影响。据报道，MiR-425-5p 参与了不同的癌症模型。然而，BM-MSCs 衍生的外泌体 miR-425-5p 在 AML 中的功能尚不清楚。
方法：通过 qRT-PCR 在临床 AML 样本中测量 miR-425-5p 的表达。使用针对 CD44、CD90 和 CD105 的抗体分析 BM-MSCs 的免疫表型。外泌体是从 BM-MSCs 中分离出来的。通过CCK-8测定、Edu测定、transwell测定、AML细胞中的流式细胞术分析BM-MSCs衍生的外泌体miR-425-5p对AML的影响。还在 AML 细胞中进行了 qRT-PCR、荧光素酶报告基因测定和蛋白质印迹分析。
结果：与健康病例的骨髓相比，原发性 AML 患者的 CD34 + CD38-AML 细胞中 miR-425-5p 的表达水平降低，而与健康病例相比，AML 患者的外泌体中 miR-425-5p 的表达水平降低。同样，与 BM-MSCs 相比，miR-425-5p 在 AML 细胞系中也下调。MiR-425-5p 能够在来自 BM-MSCs 的外泌体中表达。CCK-8、Edu、transwell 检测和流式细胞仪分析显示，BM-MSCs 衍生的外泌体 miR-425-5p 显着抑制细胞活力、Edu 阳性细胞、侵袭和迁移，并诱导 AML 细胞凋亡。同时，BM-MSCs 来源的外泌体 miR-425-5p 在细胞中增加了切割的 PARP 和切割的 caspase3 的表达水平。MiR-425-5p 抑制肾母细胞瘤 1 相关蛋白 (WTAP) 的表达。而且，
结论： BM-MSCs来源的外泌体miR-425-5p通过靶向WTAP抑制AML细胞的增殖、侵袭和迁移，诱导AML细胞凋亡。在治疗上，BM-MSCs 衍生的外泌体 miR-425-5p 可作为 AML 治疗的潜在靶点。

关键词： AML，BM-MSCs，外泌体，miR-425-5p，WTAP