Wang Jing, Li Guokai, Lin Sheng, Cheng Ling
Administration Department of Nosocomial Infection, Fujian Maternity and Child Health Hospital, No.18 Daoshan Road, Fuzhou, 350001, Fujian Province, China.
Pediatric Intensive Care Unit, Fujian Maternity and Child Health Hospital, Fuzhou, Fujian Province, China.
Biochem Genet. 2023 Feb;61(1):170-186. doi: 10.1007/s10528-022-10242-3. Epub 2022 Jul 9.
Circular RNAs (circRNAs) play important roles in human diseases, including infantile pneumonia. In this article, we aimed to investigate the functions of circ-BICC1 in lipopolysaccharide (LPS)-induced injury of WI-38 cells. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed for the levels of circ-BICC1, BICC1, microRNA-338-3p (miR-338-3p), and myeloid differentiation primary response 88 (MYD88). Cell Counting Kit-8 (CCK-8) assay, 5-ethynyl-2'-deoxyuridine (EdU) assay, and flow cytometry analysis were conducted to evaluate cell viability, proliferation, and apoptosis, respectively. Enzyme-linked immunosorbent assay (ELISA) kits were used for the concentrations of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). The levels of oxidative stress markers were detected with commercial kits. Dual-luciferase reporter assay was adopted to analyze the interaction between circ-BICC1 and miR-338-3p, as well as MYD88 and miR-338-3p. Western blot assay was employed for the protein level of MYD88. Circ-BICC1 level was increased in pneumonia patients' blood samples and LPS-treated WI-38 cells. LPS treatment suppressed WI-38 cell viability and promoted cell apoptosis, inflammation, and oxidative stress. Circ-BICC1 knockdown reversed the effect of LPS-induced WI-38 cell injury. For mechanism analysis, circ-BICC1 could function as the sponge for miR-338-3p and miR-338-3p inhibition reversed the effect of circ-BICC1 knockdown on LPS-induced WI-38 cell injury. MYD88 was identified as the target of miR-338-3p. MiR-338-3p overexpression relieved LPS-induced injury of WI-38 cells, while the impact was abolished by elevating MYD88. Circ-BICC1 silencing remitted LPS-triggered WI-38 cell damage by adsorbing miR-338-3p and regulating MYD88.
环状RNA（circRNAs）在包括婴儿肺炎在内的人类疾病中发挥着重要作用。在本文中，我们旨在研究circ-BICC1在脂多糖（LPS）诱导的WI-38细胞损伤中的功能。采用定量实时聚合酶链反应（qRT-PCR）检测circ-BICC1、BICC1、微小RNA-338-3p（miR-338-3p）和髓样分化初级反应88（MYD88）的水平。分别进行细胞计数试剂盒-8（CCK-8）检测、5-乙炔基-2'-脱氧尿苷（EdU）检测和流式细胞术分析，以评估细胞活力、增殖和凋亡。使用酶联免疫吸附测定（ELISA）试剂盒检测白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的浓度。用商业试剂盒检测氧化应激标志物的水平。采用双荧光素酶报告基因检测分析circ-BICC1与miR-338-3p以及MYD88与miR-338-3p之间的相互作用。采用蛋白质印迹法检测MYD88的蛋白水平。肺炎患者血液样本和LPS处理的WI-38细胞中circ-BICC1水平升高。LPS处理抑制WI-38细胞活力，促进细胞凋亡、炎症和氧化应激。circ-BICC1敲低可逆转LPS诱导的WI-38细胞损伤的作用。机制分析表明，circ-BICC1可作为miR-338-3p的海绵，miR-338-3p抑制可逆转circ-BICC1敲低对LPS诱导的WI-38细胞损伤的作用。MYD88被鉴定为miR-338-3p的靶标。miR-338-3p过表达可减轻LPS诱导的WI-38细胞损伤，而通过升高MYD88可消除这种影响。circ-BICC1沉默通过吸附miR-338-3p和调节MYD88减轻LPS引发的WI-38细胞损伤。
