Identification of circular RNA circVPS33A as a modulator in house dust mite-induced injury in human bronchial epithelial cells.

IF 1.5 4区医学 Q3 RESPIRATORY SYSTEM

Experimental Lung Research Pub Date : 2021-10-01 Epub Date: 2021-09-11 DOI:10.1080/01902148.2021.1974125

Yinghao Su, Limei Geng, Yunlei Ma, Xiangyan Yu, Ziyi Kang, Zenglu Kang

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引用次数: 3

Abstract

Background: House dust mite has been well documented as a major source of allergen in asthma. Circular RNAs (circRNAs) vacuolar protein sorting 33A (circVPS33A, circ_0000455) is overexpressed in a murine asthma model. Herein, we sought to identify its critical action in Dermatophagoides pteronyssinus peptidase 1 (Der p1)-induced dysfunction of BEAS-2B cells.

Methods: The levels of circVPS33A, microRNA (miR)-192-5p, and high-mobility group box 1 (HMGB1) were assessed by quantitative real-time PCR (qRT-PCR) or western blot. Actinomycin D treatment and Ribonuclease R (RNase R) assay were used to characterize circVPS33A. Cell viability, proliferation, apoptosis, migration, and invasion were evaluated by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, and transwell assays, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to quantify interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and IL-6. Direct relationship between miR-192-5p and circVPS33A or HMGB1 was verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assay.

Results: CircVPS33A was highly expressed in asthma plasma and Der p1-treated BEAS-2B cells. Knocking down circVPS33A suppressed Der p1-induced injury in BEAS-2B cells. CircVPS33A targeted miR-192-5p. MiR-192-5p directly targeted HMGB1, and miR-192-5p-mediated repression of HMGB1 alleviated Der p1-driven cell injury. Furthermore, circVPS33A modulated HMGB1 expression through miR-192-5p.

Conclusion: Our findings demonstrated that circVPS33A regulated house dust mite-induced injury in human bronchial epithelial cells at least partially depending on the modulation of the miR-192-5p/HMGB1 axis.

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环状RNA circVPS33A在尘螨诱导的人支气管上皮细胞损伤中的调节作用的鉴定

背景:尘螨是哮喘过敏原的主要来源。环状rna (circRNAs)空泡蛋白分选33A (circVPS33A, circ_0000455)在小鼠哮喘模型中过表达。在此，我们试图确定其在皮肤蜈龙翼窦肽酶1 (Der p1)诱导的BEAS-2B细胞功能障碍中的关键作用。方法:采用实时荧光定量PCR (qRT-PCR)或western blot检测circVPS33A、microRNA (miR)-192-5p、高迁移率组盒1 (HMGB1)的表达水平。采用放线菌素D处理和核糖核酸酶R (RNase R)检测对circVPS33A进行表征。分别采用细胞计数试剂盒-8 (CCK-8)、5-乙基-2′-脱氧尿苷(EdU)、流式细胞术和transwell法评估细胞活力、增殖、凋亡、迁移和侵袭。采用酶联免疫吸附法(ELISA)定量白细胞介素-1β (IL-1β)、肿瘤坏死因子-α (TNF-α)、白细胞介素-6。通过双荧光素酶报告基因和RNA免疫沉淀(RIP)实验验证miR-192-5p与circVPS33A或HMGB1之间的直接关系。结果:CircVPS33A在哮喘血浆和Der p1处理的BEAS-2B细胞中高表达。敲除circVPS33A可抑制Der p1诱导的BEAS-2B细胞损伤。CircVPS33A靶向miR-192-5p。MiR-192-5p直接靶向HMGB1, MiR-192-5p介导的HMGB1抑制减轻了Der p1驱动的细胞损伤。此外，circVPS33A通过miR-192-5p调节HMGB1的表达。结论:我们的研究结果表明，circVPS33A至少部分地通过miR-192-5p/HMGB1轴的调节来调节尘螨诱导的人支气管上皮细胞损伤。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Experimental Lung Research 医学-呼吸系统

CiteScore

3.80

自引率

0.00%

发文量

审稿时长

2 months

期刊介绍： Experimental Lung Research publishes original articles in all fields of respiratory tract anatomy, biology, developmental biology, toxicology, and pathology. Emphasis is placed on investigations concerned with molecular, biochemical, and cellular mechanisms of normal function, pathogenesis, and responses to injury. The journal publishes reports on important methodological advances on new experimental modes. Also published are invited reviews on important and timely research advances, as well as proceedings of specialized symposia. Authors can choose to publish gold open access in this journal.