Joint Surgery and Sports Medicine Department, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, China.
J Gene Med. 2021 Feb;23(2):e3304. doi: 10.1002/jgm.3304. Epub 2021 Jan 20.
Growing evidence suggests that circular RNAs (circRNAs) are involved in the development of osteoarthritis (OA). The present study aimed to explore the CircADAMTS6/miR-431-5p axis with respect to regulating interleukin-1β (IL-1β) induced chondrocyte apoptosis.
We first evaluated the differentially expressed circRNAs between normal chondrocytes and interleukin (IL)-1β-stimulated chondrocytes. Then, bioinformatic analysis was performed to identify the role and function of circADAMTS6. Small interfering RNA-expressing or overexpressing circADAMTS6 lentiviral vectors were used for transduction of chondrocytes. Annexin-V-fluorescein isothiocyanate (FITC) double staining was performed to measure the apoptotic rate of the chondrocytes in each group. Finally, a dual luciferase reporter assay was performed to identify the target relationship between circADAMTS6 and miR-431-5p.
After treatment with IL-1β, circADAMTS6 was down-regulated compared to the normal chondrocyte group. The overexpression of circADAMTS6 inhibited apoptosis in human chondrocytes, as indicated by annexin-V-FITC double staining. However, overexpression of miR-431-5p had the opposite effect. A dual luciferase reporter assay indicated that circADAMTS6 could directly binding with miR-431-5p.
Our findings demonstrate that the circADAMTS6/miR-431-5p axis comprises a new target for OA. Bioinformatic analysis suggested that circADAMTS6 acted as a sponge of miR-431-5p.
越来越多的证据表明,环状 RNA(circRNA)参与了骨关节炎(OA)的发生。本研究旨在探讨 CircADAMTS6/miR-431-5p 轴在调节白细胞介素-1β(IL-1β)诱导的软骨细胞凋亡中的作用。
首先评估正常软骨细胞和白细胞介素(IL)-1β刺激的软骨细胞之间差异表达的 circRNAs。然后,进行生物信息学分析以确定 CircADAMTS6 的作用和功能。用表达小干扰 RNA 的或过表达 CircADAMTS6 的慢病毒载体转导软骨细胞。用 Annexin-V-异硫氰酸荧光素(FITC)双重染色法测量各组软骨细胞的凋亡率。最后,进行双荧光素酶报告基因检测以鉴定 CircADAMTS6 和 miR-431-5p 之间的靶关系。
与正常软骨细胞组相比,IL-1β 处理后 CircADAMTS6 下调。CircADAMTS6 的过表达通过 Annexin-V-FITC 双重染色抑制人软骨细胞凋亡。然而,miR-431-5p 的过表达则有相反的效果。双荧光素酶报告基因检测表明 CircADAMTS6 可以直接与 miR-431-5p 结合。
我们的研究结果表明,CircADAMTS6/miR-431-5p 轴构成了 OA 的一个新靶点。生物信息学分析表明 CircADAMTS6 作为 miR-431-5p 的海绵。