基因敲除
阿格里坎
软骨细胞
免疫印迹
血管内皮生长因子A
小RNA
细胞凋亡
化学
流式细胞术
分子生物学
骨关节炎
癌症研究
血管内皮生长因子
生物
体外
医学
血管内皮生长因子受体
基因
病理
生物化学
关节软骨
替代医学
作者
Wei Xie,Luoyong Jiang,Xiaoyang Huang,Wei You,Wei Sun
出处
期刊:Current Molecular Medicine
[Bentham Science]
日期:2024-02-01
卷期号:24 (2): 217-225
被引量:2
标识
DOI:10.2174/1566524023666221103161203
摘要
Objective: Circular RNAs (circRNAs) have been extensively implicated in osteoarthritis (OA) progression. Therefore, this study explores the impact of hsa_circ_00046621 on OA progression and the related molecular mechanism. Methods: Human articular chondrocyte injury was induced by IL-1β to construct the OA model in vitro. hsa_circ_0004662 and microRNA (miR)-424-5p expression in chondrocytes was evaluated with qRT-PCR. Vascular endothelial growth factors A (VEGFA) expression was examined with qRT-PCR and western blot after hsa_circ_0004662 knockdown or miR-424-5p overexpression in chondrocytes. Subsequent to loss- and gain-of-function assays in IL-1β-induced chondrocytes, the proliferation and apoptosis of chondrocytes were assessed with CCK-8 assay and flow cytometry, respectively. The expression of MMP13, Aggrecan, and apoptosis-related proteins Bax and Bcl-2 was measured with western blot. The binding of miR-424-5p to hsa_circ_0004662 and VEGFA was assessed with a dual-luciferase reporter gene assay. Results: Hsa_circ_0004662 was up-regulated, but miR-424-5p was down-regulated in IL-1β-induced chondrocytes. Mechanistically, both hsa_circ_0004662 and VEGFA bound to miR-424-5p, and hsa_circ_0004662 enhanced VEGFA expression by down-regulating miR-424-5p. Hsa_circ_0004662 knockdown elevated cell proliferation, decreased apoptosis and MMP13 and Bax expression, and increased Aggrecan and Bcl-2 expression in IL-1β-induced chondrocytes, which was counteracted by further miR-424-5p down-regulation or VEGFA overexpression. method: Human articular chondrocyte injury was induced by IL-1β to mimic the OA model. hsa_circ_0004662 and microRNA (miR)-424-5p expression in chondrocytes was evaluated using qRT-PCR. Vascular endothelial growth factors A (VEGFA) expression was examined using qRT-PCR and western blot analysis following hsa_circ_0004662 knockdown or miR-424-5p overexpression in chondrocytes. Subsequent to the loss- and gain-of-function assays in IL-1β-induced chondrocytes, proliferation and apoptosis of chondrocytes were assessed using CCK-8 assay and flow cytometry, respectively. The expression of MMP13, Aggrecan, and apoptosis-related proteins Bax and Bcl-2 was measured using western blot analysis. The binding relationship of miR-424-5p to hsa_circ_0004662 and VEGFA was assessed using dual luciferase reporter gene assay. Conclusion: Hsa_circ_0004662 facilitates OA progression via the miR-424-5p/VEGFA axis.
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