Orthopaedic Surgery, Renmin Hospital of Wuhan University, Wuhan, Hubei, China.
Aging (Albany NY). 2020 Nov 6;12(21):21971-21991. doi: 10.18632/aging.104035.
The purpose of this study was to identify a specific circular RNA and to investigate its regulatory mechanism in intervertebral disc degeneration (IDD). CircGLCE was selected after microarray analyses and was further analysed by RT-qPCR and FISH. CircGLCE was found to stably exist in the cytoplasm of nucleus pulposus (NP) cells. It was downregulated in IDD. After silencing CircGLCE, its function was assessed with RT-qPCR, immunofluorescence analysis and flow cytometry. Knockdown of CircGLCE promoted apoptosis and induced the expression of matrix-degrading enzymes in NP cells. CircGLCE served as a miR-587 sponge in NP cells. Inhibiting miR-587 counteracted the IDD-enhancing effect caused by silencing CircGLCE. STAP1 served as the miRNA target that mediated the functions of miR-587. In an IDD mouse model, the in vivo effects of overexpressing CircGLCE on IDD were confirmed by imaging techniques, TUNEL staining, FISH, western blotting, H&E staining and immunohistochemistry. Thus, CircGLCE attenuates IDD by inhibiting the apoptosis of NP cells and ECM degradation through the targeting of miR-587/STAP1. CircGLCE may be a potential therapeutic target for IDD treatments.
本研究旨在鉴定一种特定的环状 RNA,并研究其在椎间盘退变 (IDD) 中的调控机制。通过微阵列分析选择 CircGLCE,并通过 RT-qPCR 和 FISH 进一步分析。发现 CircGLCE 稳定存在于核髓核 (NP) 细胞的细胞质中。在 IDD 中下调。沉默 CircGLCE 后,通过 RT-qPCR、免疫荧光分析和流式细胞术评估其功能。沉默 CircGLCE 可促进 NP 细胞凋亡并诱导基质降解酶的表达。CircGLCE 在 NP 细胞中作为 miR-587 的海绵。在 NP 细胞中,抑制 miR-587 可逆转沉默 CircGLCE 引起的 IDD 增强作用。STAP1 作为介导 miR-587 功能的 miRNA 靶标。在 IDD 小鼠模型中,通过成像技术、TUNEL 染色、FISH、western blot、H&E 染色和免疫组织化学证实过表达 CircGLCE 对 IDD 的体内作用。因此,CircGLCE 通过靶向 miR-587/STAP1 抑制 NP 细胞凋亡和 ECM 降解来减轻 IDD。CircGLCE 可能是治疗 IDD 的潜在治疗靶点。