Circ-EC1可能与miR-708-5p竞争调控支气管肺发育不良中的Ntrk2

IF 3 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Hanrong Cheng, Dongcai Li, Yuming Tang, Tianyong Hu, Benqing Wu
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引用次数: 0

摘要

支气管肺发育不良(BPD)影响患者的生活质量。环状 RNA 参与了 BPD 的研究。然而,circ-ECH1在BPD中的作用尚未见报道。本研究旨在探讨 circ-ECH1 在 BPD 中的作用和机制。研究采用高氧处理的 II 型肺泡上皮细胞(L2 细胞)作为体外 BPD 模型。使用 CCK-8、流式细胞术和活性氧(ROS)评估细胞活力。荧光原位杂交证实了亚细胞定位。研究人员使用circ-ECH1过表达(或抑制)和miR-708-5p模拟物来研究circ-ECH1和miR-708-5p在BPD中的作用。定量反转录聚合酶反应(qRT-PCR)检测了circ-EC1、miR-708-5p和神经营养受体酪氨酸激酶2(Ntrk2)的表达。通过 Western 印迹分析评估了 Ntrk2 的表达。通过苏木精和伊红染色评估肺组织的变化。肺纤维化通过 Mason 染色法进行检测。TUNEL染色用于评估肺组织中的细胞凋亡。对 BPD 大鼠的肺组织进行了 RNA 测序。通过双重荧光素酶活性证实了 circ-ECH1 与 miR-708-5p 之间的结合。高氧降低了细胞活力,增加了细胞凋亡和 ROS 积累。此外,高氧还降低了主要位于细胞质中的 circ-ECH1 的表达水平。Circ-ECH1 的过表达提高了细胞活力,但减少了细胞凋亡和 ROS 积累。相反,干扰 circ-ECH1 会进一步促进细胞凋亡并降低细胞活性。此外,circ-EC1 的过表达还降低了肺纤维化和肺细胞凋亡的发生率。RNA 测序和 qRT-PCR 证实,Ntrk2 和 miR-708-5p 的表达受 circ-ECH1 的影响。从机理上讲,circ-EC1 可能与 miR-708-5p 结合以调控 Ntrk2 的表达。circ-ECH1 可能与 miR-708-5p 竞争,从而调节 Ntrk2 在 BPD 中的表达。这些发现为治疗 BPD 提供了一个新的靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Circ-ECH1 May Compete With miR-708-5p to Regulate Ntrk2 in Bronchopulmonary Dysplasia.

Bronchopulmonary dysplasia (BPD) affects patients' quality of life. Circular RNAs participated in BPD. However, circ-ECH1's role in BPD has not been reported yet. This study aimed to explore the role and mechanism of circ-ECH1 in BPD. Hyperoxia-treated type II alveolar epithelial cells (L2 cells) were used as the in vitro BPD model. CCK-8, flow cytometry, and reactive oxygen species (ROS) were used to evaluate cell viability. Fluorescence in situ hybridization confirmed the subcellular localization. Circ-ECH1 overexpression (or inhibited) and miR-708-5p mimics were used to investigate the roles of circ-ECH1 and miR-708-5p in BPD. Quantitative reverse-transcription polymerase reaction (qRT-PCR) detected the expressions of circ-ECH1, miR-708-5p, and neurotrophic receptor tyrosine kinase 2 (Ntrk2). Ntrk2 expression was evaluated by Western blot analysis. Changes in lung tissues were evaluated by hematoxylin and eosin staining. Pulmonary fibrosis was examined by Mason staining. TUNEL staining was performed to evaluate cell apoptosis in lung tissues. RNA sequencing was performed in the lung tissues of BPD rats. The binding between circ-ECH1 and miR-708-5p was confirmed through dual luciferase activity. Hyperoxia reduced cell viability and increased cell apoptosis and ROS accumulation. In addition, hyperoxia decreased the expression levels of circ-ECH1, which is mainly located in the cytoplasm. Circ-ECH1 overexpression increased cell viability but reduced cell apoptosis and ROS accumulation. On the contrary, interference with circ-ECH1 further promoted cell apoptosis and reduced cell activity. Furthermore, circ-ECH1 overexpression reduced the incidence of pulmonary fibrosis and lung cell apoptosis. RNA sequencing, followed by qRT-PCR, confirmed that the expressions of Ntrk2 and miR-708-5p were affected by circ-ECH1. miR-708-5p mimics reversed the role of circ-ECH1 in the BPD. Mechanistically, circ-ECH1 may bind with miR-708-5p to regulate Ntrk2 expression. Circ-ECH1 may compet with miR-708-5p to regulate Ntrk2 expression in BPD. The findings provided a new target for BPD treatment.

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来源期刊
Journal of cellular biochemistry
Journal of cellular biochemistry 生物-生化与分子生物学
CiteScore
9.90
自引率
0.00%
发文量
164
审稿时长
1 months
期刊介绍: The Journal of Cellular Biochemistry publishes descriptions of original research in which complex cellular, pathogenic, clinical, or animal model systems are studied by biochemical, molecular, genetic, epigenetic or quantitative ultrastructural approaches. Submission of papers reporting genomic, proteomic, bioinformatics and systems biology approaches to identify and characterize parameters of biological control in a cellular context are encouraged. The areas covered include, but are not restricted to, conditions, agents, regulatory networks, or differentiation states that influence structure, cell cycle & growth control, structure-function relationships.
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