Department of Pediatrics, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, No.321. Zhongshan Road, 210008 Nanjing, Jiangsu, China; Department of Pediatrics, The Fourth Affiliated Hospital of Nanjing Medical University, No. 298, Nanpu Road, 210031 Nanjing, Jiangsu, China.
Department of Pediatrics, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, No.321. Zhongshan Road, 210008 Nanjing, Jiangsu, China.
Cell Signal. 2022 Jul;95:110340. doi: 10.1016/j.cellsig.2022.110340. Epub 2022 Apr 26.
Excessive proliferation and migration of airway smooth muscle cells (ASMCs) directly lead to airway remodeling in asthma. However, the role of circular RNAs (circRNAs) in airway remodeling remains unclear. This study aimed to investigate the regulatory role and mechanism of circ_CSNK1E in ASMCs proliferation and migration.
In this study, RNA-sequencing was used to analyze cicRNAs expression in asthma samples. ASMCs were treated with 25 ng/ml PDGF-BB to establish a model of asthma in vitro. Then, we used RT-qPCR to assess circRNAs, microRNAs (miRNAs) and messenger RNAs (mRNAs) expression. Besides, CCK-8, colony formation, wound healing and transwell chamber assays were carried out to explore cell proliferation and migration. Subcellular localization assay was used to detect the location of circRNA. Next, bioinformatics, luciferase reporter and RIP assays were performed to evaluate the relationship among circ_CSNK1E, miRNA-34a-5p and VAMP2.
circ_CSNK1E expression was found to be significantly up-regulated in asthma samples and PDGF-BB-induced ASMCs. Functional experiments revealed that inhibition of circRNA_CSNK1E suppressed proliferation and migration of ASMCs stimulated by PDGF-BB. Next, we found that circRNA_CSNK1E served as a sponge for miR-34a-5p in ASMCs, and miR-34a-5p mimic suppressed proliferation and migration of ASMCs. Moreover, VAMP2 was confirmed as a direct target of miR-34a-5p. At last, inhibition of circRNA_CSNK1E suppressed proliferation and migration of ASMCs stimulated by PDGF-BB through miR-34a-5p/VAMP2 axis.
Collectively, these findings clarified the importance of circ_CSNK1E/miRNA-34a-5p/VAMP2 axis for the proliferation and migration of ASMCs. These indicated that inhibition of circ_CSNK1E might be a potential target for the treatment of airway remodeling in asthma.
气道平滑肌细胞（ASMCs）的过度增殖和迁移直接导致哮喘中的气道重塑。然而，环状 RNA（circRNAs）在气道重塑中的作用尚不清楚。本研究旨在探讨 circ_CSNK1E 在 ASMCs 增殖和迁移中的调控作用和机制。
本研究采用 RNA 测序分析哮喘样本中 circRNAs 的表达。用 25ng/ml PDGF-BB 处理 ASMCs 建立体外哮喘模型。然后，我们用 RT-qPCR 评估 circRNAs、microRNAs（miRNAs）和信使 RNA（mRNAs）的表达。此外，进行 CCK-8、集落形成、划痕愈合和 Transwell 室分析以探索细胞增殖和迁移。亚细胞定位测定用于检测 circRNA 的位置。接下来，进行生物信息学、荧光素酶报告和 RIP 测定以评估 circ_CSNK1E、miRNA-34a-5p 和 VAMP2 之间的关系。
circ_CSNK1E 的表达在哮喘样本和 PDGF-BB 诱导的 ASMCs 中明显上调。功能实验表明，抑制 circRNA_CSNK1E 可抑制 PDGF-BB 刺激的 ASMCs 的增殖和迁移。接下来，我们发现 circRNA_CSNK1E 可作为 ASMCs 中的 miR-34a-5p 海绵，并且 miR-34a-5p 模拟物可抑制 ASMCs 的增殖和迁移。此外，VAMP2 被证实是 miR-34a-5p 的直接靶标。最后，抑制 circRNA_CSNK1E 通过 miR-34a-5p/VAMP2 轴抑制 PDGF-BB 刺激的 ASMCs 的增殖和迁移。
总之，这些发现阐明了 circ_CSNK1E/miRNA-34a-5p/VAMP2 轴对 ASMCs 增殖和迁移的重要性。这表明抑制 circ_CSNK1E 可能是治疗哮喘气道重塑的潜在靶点。
