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miR-486-5p 通过靶向 FOXO1 抑制白血病细胞增殖并诱导细胞凋亡。

MiR-486-5p inhibits the proliferation of leukemia cells and induces apoptosis through targeting FOXO1.

机构信息

Department of Hematology, The First Affiliated Hospital of Xi'an Medical University, China.

Department of Hematology, The First Affiliated Hospital of Xi'an Medical University, China.

出版信息

Mol Cell Probes. 2019 Apr;44:37-43. doi: 10.1016/j.mcp.2019.02.001. Epub 2019 Feb 5.

Abstract

AIM

Studies have reported that micro (miR)-486-5p plays a crucial part in the progression of leukemia, however, to the best of our knowledge, few studies have been conducted on its mechanism in leukemia. In this study, the mechanism of miR-486-5p in leukemia cells was pointed out and its possible target genes were analyzed for the purpose of providing new therapeutic strategies for treating leukemia patients.

METHODS

MiRNA expression of Leukemia cells (K562, Kasumi-1, and THP-1) and primary leukocytes was detected by Real-time Quantitative polymerase chain reaction(qPCR). The activity of the cells was assessed using the Cell Counting Kit-8 (CCK-8). Apoptotic cells were analyzed by a flow cytometer (FCM). Caspase-3 activation in leukemia cells was determined by Western blot. Targetscan 7.2 was used to predict the potential targets of miR-486-5p and further confirmed by dual-luciferase reporter assay.

RESULT

miR-486-5p was significantly down-regulated in leukemia cells. The over-expression of miR-486-5p notably increased the apoptosis and caspase-3 activity in leukemia cells. There was a predicted interaction site for miR-486-5p in the FOXO1 3'-UTR. Furthermore, this study showed that FOXO1 was significantly up-regulated in leukemia cells, the growth of which was depressed by the up-regulation of miR-486-5p.

CONCLUSION

miR-486-5p may inhibit the proliferation of leukemia cells and induce apoptosis through targeting FOXO1.

摘要

目的

已有研究报道 miR-486-5p 在白血病的进展中发挥着关键作用,但就我们所知,针对其在白血病中的作用机制的研究甚少。本研究旨在指出 miR-486-5p 在白血病细胞中的作用机制,并分析其可能的靶基因,以期为白血病患者的治疗提供新的策略。

方法

采用实时定量聚合酶链反应(qPCR)检测白血病细胞(K562、Kasumi-1 和 THP-1)和原代白细胞中的 miRNA 表达。采用细胞计数试剂盒-8(CCK-8)评估细胞活性。采用流式细胞仪(FCM)分析凋亡细胞。采用 Western blot 检测白血病细胞中 caspase-3 的激活。利用 Targetscan 7.2 预测 miR-486-5p 的潜在靶基因,并通过双荧光素酶报告基因实验进一步验证。

结果

miR-486-5p 在白血病细胞中表达明显下调。miR-486-5p 的过表达显著增加白血病细胞的凋亡和 caspase-3 活性。在 FOXO1 3'-UTR 中存在 miR-486-5p 的预测作用靶点。此外,本研究表明 FOXO1 在白血病细胞中明显上调,miR-486-5p 的上调抑制了其生长。

结论

miR-486-5p 可能通过靶向 FOXO1 抑制白血病细胞的增殖并诱导其凋亡。

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