Analysis of potential functional significance of microRNA-3613-3p in human umbilical vein endothelial cells affected by heat stress

Abstract

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-quan¬ titative 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. M A P 3 K 2 protein was demonstrated to be upregu-lated in heat-treated Ecs, and overexpression of miR-3613-3p reduced M A P 3 K 2 expression levels. additionally, M A P 3 K 2 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.