Dai Jing, Wang Kai, Liu Tao, Wang Qiong, Pang Yingxu
Department of Hematology, Zhoukou Central Hospital No. 26, Renmin East Road, Zhoukou 466000 Zhengzhou China
RSC Adv. 2019 Nov 7;9(62):36256-36265. doi: 10.1039/c9ra06378a. eCollection 2019 Nov 4.
Recent studies show that lncRNA KCNQ1OT1 and microRNA-186-5p (miR-186-5p) are involved in various human cancers. Moreover, it is reported that KCNQ1OT1 expression is upregulated in acute myeloid leukemia (AML). However, their roles in AML remain unknown. This study aimed to reveal the functional mechanism of KCNQ1OT1 and miR-186-5p in AML development. Quantitative real time polymerase chain reaction (qRT-PCR) was performed to detect the levels of genes. Cell proliferation and apoptosis were assessed by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2--tetrazolium bromide (MTT) assay and flow cytometry analysis respectively. A western blot assay was carried out to examine the protein levels. In addition, the interaction between miR-186-5p and KCNQ1OT1 or neural cell adhesion molecule 1 (NCAM1) was predicted by bioinformatics analysis tool starbase2.0 and confirmed by the dual luciferase reporter assay. KCNQ1OT1 and NCAM1 expressions were increased and miR-186-5p expression was decreased in AML samples and cells. The depletion of KCNQ1OT1 inhibited cell proliferation, and promoted apoptosis and chemo-sensitivity in AML. In addition, the upregulation of miR-186-5p suppressed AML cell proliferation, and induced apoptosis and chemo-sensitivity. Interestingly, KCNQ1OT1 directly downregulated miR-186-5p expression and miR-186-5p decreased NCAM1 expression by binding to the 3' untranslated region (UTR) of NCAM1 mRNA. Furthermore, miR-186-5p knockdown or NCAM1 overexpression reversed the effects of KCNQ1OT1 depletion on AML cell progression. Our results firstly revealed a linear relationship between KCNQ1OT1, miR-186-5p, and NCAM1, and demonstrated that KCNQ1OT1 mediated AML cell progression regulating the miR-186-5p/NCAM1 axis, revealing functional mechanisms of KCNQ1OT1 and miR-186-5p in AML development.
最近的研究表明，长链非编码RNA KCNQ1OT1和微小RNA-186-5p（miR-186-5p）参与了多种人类癌症。此外，据报道KCNQ1OT1在急性髓系白血病（AML）中表达上调。然而，它们在AML中的作用仍不清楚。本研究旨在揭示KCNQ1OT1和miR-186-5p在AML发生发展中的功能机制。采用定量实时聚合酶链反应（qRT-PCR）检测基因水平。分别通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-溴化四氮唑（MTT）法和流式细胞术分析评估细胞增殖和凋亡。进行蛋白质印迹分析以检测蛋白质水平。此外，通过生物信息学分析工具starbase2.0预测miR-186-5p与KCNQ1OT1或神经细胞黏附分子1（NCAM1）之间的相互作用，并通过双荧光素酶报告基因检测进行验证。在AML样本和细胞中，KCNQ1OT1和NCAM1表达增加，而miR-186-5p表达降低。KCNQ1OT1的缺失抑制了AML细胞增殖，并促进了细胞凋亡和化疗敏感性。此外，miR-186-5p的上调抑制了AML细胞增殖，并诱导了细胞凋亡和化疗敏感性。有趣的是，KCNQ1OT1直接下调miR-186-5p的表达，而miR-186-5p通过与NCAM1 mRNA的3'非翻译区（UTR）结合降低NCAM1的表达。此外，miR-186-5p的敲低或NCAM1的过表达逆转了KCNQ1OT1缺失对AML细胞进程的影响。我们的结果首次揭示了KCNQ1OT1、miR-186-5p和NCAM1之间的线性关系，并证明KCNQ1OT1通过调节miR-186-5p/NCAM1轴介导AML细胞进程，揭示了KCNQ1OT1和miR-186-5p在AML发生发展中的功能机制。
