Department of Emergency, Central Theater General Hospital of People's Liberation Army of China, Wuhan, Hubei 430070, P.R. China.
Department of General Surgery, Central Theater General Hospital of People's Liberation Army of China, Wuhan, Hubei 430070, P.R. China.
Mol Med Rep. 2019 Aug;20(2):1846-1856. doi: 10.3892/mmr.2019.10376. Epub 2019 Jun 11.
Dysregulation of microRNA‑3613‑3p (miR‑3613‑3p) was previously reported in endothelial cells (ECs) during heat stress. The aim of the present study was to investigate the precise role of miR‑3613‑3p in heat stress. In the present study, potential gene targets of miR‑3613‑3p in heat‑treated ECs were assessed, and the potential effects of miR‑3613‑3p were determined using Gene Ontology enrichment analysis. Kyoto Encyclopedia of Genes and Genomes pathway analysis was used to identify signaling pathways that may be affected by miR‑3613‑3p in heat‑treated cells. Reverse transcription‑quantitative PCR, western blotting and annexin V‑FITC/propidium iodide staining were performed to detect miRNA expression, protein expression and apoptosis, respectively. Luciferase gene reporter assay was performed to evaluate the association between miR‑3613‑3p and mitogen‑activated protein kinase kinase kinase 2 (MAP3K2). Bioinformatics analysis revealed 865 potential gene targets for miR‑3613‑3p and a series of functions and pathways in heat‑treated ECs. 'Negative regulation of apoptotic process' was identified as a potential function of miR‑3613‑3p. In addition, functional analysis confirmed the downregulated expression levels of miR‑3613‑3p in ECs during heat stress, which was accompanied by an increase in apoptosis; restoration of miR‑3613‑3p expression inhibited apoptosis. MAP3K2 protein was demonstrated to be upregulated in heat‑treated ECs, and overexpression of miR‑3613‑3p reduced MAP3K2 expression levels. Additionally, MAP3K2 was targeted by miR‑3613‑3p. These results indicated that miR‑3613‑3p may have complicated roles in ECs under heat stress. miR‑3613‑3p may serve an important role in the apoptosis of heat‑treated ECs, and this effect may be partly achieved by targeting MAP3K2.
miR-3613-3p 的失调先前在热应激过程中的内皮细胞(ECs)中被报道。本研究的目的是探究 miR-3613-3p 在热应激中的精确作用。在本研究中,评估了 miR-3613-3p 在热处理的 ECs 中的潜在基因靶标,并通过基因本体论富集分析确定了 miR-3613-3p 的潜在影响。京都基因与基因组百科全书通路分析用于鉴定可能受 miR-3613-3p 影响的热处理细胞中的信号通路。逆转录-定量 PCR、western blot 和 Annexin V-FITC/碘化丙啶染色分别用于检测 miRNA 表达、蛋白表达和细胞凋亡。荧光素酶基因报告实验用于评估 miR-3613-3p 与丝裂原活化蛋白激酶激酶激酶 2(MAP3K2)之间的关联。生物信息学分析显示,miR-3613-3p 有 865 个潜在的基因靶标,以及一系列在热处理的 ECs 中的功能和通路。“凋亡过程的负调控”被鉴定为 miR-3613-3p 的潜在功能。此外,功能分析证实了 miR-3613-3p 在热应激过程中的下调表达水平,伴随着细胞凋亡的增加;miR-3613-3p 表达的恢复抑制了细胞凋亡。MAP3K2 蛋白在热处理的 ECs 中被上调,并且 miR-3613-3p 的过表达降低了 MAP3K2 的表达水平。此外,MAP3K2 是 miR-3613-3p 的靶标。这些结果表明,miR-3613-3p 在热应激下的 ECs 中可能具有复杂的作用。miR-3613-3p 可能在热处理的 ECs 凋亡中发挥重要作用,这种作用可能部分通过靶向 MAP3K2 来实现。