Jiang Chen, Xu Ming, Zhu Jin, Yang Dongrong, Xue Boxin
Department of Urology, The Second Affiliated Hospital of Soochow University, Suzhou, China.
Drug Dev Res. 2022 Apr;83(2):351-361. doi: 10.1002/ddr.21864. Epub 2021 Aug 9.
Circular RNA (circRNA) have been found to play an important role in the progression of many diseases, including interstitial cystitis (IC). However, the role of circTHBS1 in IC progression is still unclear. Exploring the role and potential molecular mechanism of circTHBS1 in the development of IC. The enzyme-linked immunosorbent assay was used to assess the levels of inflammatory cytokines. The expression levels of circTHBS1, microRNA (miR)-139-5p, and mitofusin 2 (MFN2) were evaluated using quantitative real-time PCR. Cell proliferation and migration were determined using MTT assay, Edu staining, and transwell assay. The protein levels of epithelial-mesenchymal transition (EMT) markers and MFN2 were examined using western blot analysis. The relationship between miR-139-5p and circTHBS1 or MFN2 was confirmed using the dual-luciferase reporter assay and RIP assay. CircTHBS1 was highly repressed in IC tissues and cells, and its expression was positively correlated with the inflammatory response of IC patients. CircTHBS1 could promote the proliferation, migration, EMT process, and inflammation of IC cells, while its knockdown had an opposite effect. CircTHBS1 could serve as a sponge of miR-139-5p, and miR-139-5p could participate in the regulation of circTHBS1 on IC cell progression. In addition, miR-139-5p could target MFN2, and it could inhibit the progression of IC cells by targeting MFN2. Furthermore, circTHBS1 sponged miR-139-5p to positively regulate MFN2. CircTHBS1 promoted IC cell proliferation, migration, EMT process, and inflammation by regulating the miR-139-5p/MFN2 axis indicating that circTHBS1 might be a potential target for IC treatment.
环状RNA（circRNA）已被发现在包括间质性膀胱炎（IC）在内的多种疾病进展中发挥重要作用。然而，circTHBS1在IC进展中的作用仍不清楚。本研究旨在探讨circTHBS1在IC发生发展中的作用及潜在分子机制。采用酶联免疫吸附测定法评估炎性细胞因子水平。运用定量实时PCR评估circTHBS1、微小RNA（miR）-139-5p和线粒体融合蛋白2（MFN2）的表达水平。采用MTT法、EdU染色和Transwell实验检测细胞增殖和迁移情况。运用蛋白质免疫印迹分析检测上皮-间质转化（EMT）标志物和MFN2的蛋白水平。通过双荧光素酶报告基因实验和RNA免疫沉淀实验验证miR-139-5p与circTHBS1或MFN2之间的关系。circTHBS1在IC组织和细胞中表达显著下调，其表达与IC患者的炎症反应呈正相关。circTHBS1可促进IC细胞的增殖、迁移、EMT进程和炎症反应，而敲低circTHBS1则产生相反作用。circTHBS1可作为miR-139-5p的海绵，且miR-139-5p可参与circTHBS1对IC细胞进程的调控。此外，miR-139-5p可靶向MFN2，通过靶向MFN2抑制IC细胞进程。此外，circTHBS1通过吸附miR-139-5p正向调控MFN2。circTHBS1通过调控miR-139-5p/MFN2轴促进IC细胞增殖、迁移、EMT进程和炎症反应，表明circTHBS1可能是IC治疗的潜在靶点。
