Department of Orthopaedic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine; No.88 Jiefang Road, Hangzhou 310009, P.R. China.
Curr Gene Ther. 2022;22(5):427-438. doi: 10.2174/1566523222666220309150722.
Osteoarthritis (OA) is the predominant threat to the health of the elderly, and it is crucial to understand the molecular pathogenetic mechanisms involved in it. This study aims to investigate the role of a well-studied cancer-related long non-coding RNA (lncRNA)-POU3F3 in OA and its implicated molecular mechanisms.
The expression of POU3F3 and miR-29a-3p was examined in osteoarthritis patients, as well as destabilization of the medial meniscus (DMM) mouse OA model and IL- 1β induced chondrocytes cell OA model, by quantitative real-time PCR. The interaction between POU3F3, miR-29a-3p and transcription factor forkhead box O3 (FOXO3) was verified via dual-luciferase reporter analysis and RNA immunoprecipitation analyses. Cell proliferation and apoptosis were evaluated by cell viability assay and flow cytometry, respectively. Cartilage extracellular matrix (ECM) degradation was investigated with ELISA and western blotting. In addition, the in vivo regulation of POU3F3 in OA was verified by intra-articular injection of lentivirus overexpression POU3F31 in mice models.
The expression level of POU3F3 was decreased in OA patients/animal cartilage tissues and IL-1β-stimulated in vitro chondrocyte model. POU3F3 overexpression inhibited IL-1β-induced injury of chondrocytes, enhancing cell viability, suppressing apoptosis and inflammatory cytokine secretion, rescuing metabolic dysfunction, and restraining autophagy in vitro. Mechanistically, Luciferase reporter and RNA immunoprecipitation (RIP) assays indicated that miR-29a-3p could directly bind to POU3F3, and FOXO3 was a target gene of miR-29a-3p. Functional rescue assays confirmed this POU3F3/miR-29a-3p/FOXO3 axis in chondrocytes during OA occurrence. Furthermore, intraarticularly delivery of lentivirus containing POU3F3 alleviates the damage in mouse OA model in vivo.
In conclusion, this work highlights the role of the POU3F3/miR-29a-3p/FOXO3 axis in the OA pathogenesis, suggesting this axis as a potential therapeutic target for OA.
骨关节炎（OA）是老年人健康的主要威胁，了解其涉及的分子发病机制至关重要。本研究旨在探讨一种研究充分的癌症相关长链非编码 RNA（lncRNA）-POU3F3 在 OA 中的作用及其潜在的分子机制。
通过定量实时 PCR 检测 OA 患者、内侧半月板不稳定（DMM）小鼠 OA 模型和 IL-1β诱导的软骨细胞 OA 模型中 POU3F3 和 miR-29a-3p 的表达。通过双荧光素酶报告分析和 RNA 免疫沉淀分析验证 POU3F3、miR-29a-3p 和转录因子叉头框 O3（FOXO3）之间的相互作用。通过细胞活力测定和流式细胞术分别评估细胞增殖和细胞凋亡。通过 ELISA 和 Western blot 分析检测软骨细胞外基质（ECM）降解。此外，通过向小鼠模型关节内注射过表达 POU3F3 的慢病毒来验证 POU3F3 在 OA 中的体内调节作用。
在 OA 患者/动物软骨组织和 IL-1β刺激的体外软骨细胞模型中，POU3F3 的表达水平降低。POU3F3 过表达抑制了 IL-1β诱导的软骨细胞损伤，增加了细胞活力，抑制了细胞凋亡和炎症细胞因子的分泌，恢复了体外代谢功能障碍，并抑制了自噬。机制上，荧光素酶报告和 RNA 免疫沉淀（RIP）试验表明，miR-29a-3p 可以直接与 POU3F3 结合，而 FOXO3 是 miR-29a-3p 的靶基因。功能挽救试验证实了 OA 发生过程中软骨细胞中的 POU3F3/miR-29a-3p/FOXO3 轴。此外，关节内递送含有 POU3F3 的慢病毒在体内减轻了小鼠 OA 模型的损伤。
总之，这项工作强调了 POU3F3/miR-29a-3p/FOXO3 轴在 OA 发病机制中的作用，表明该轴可能成为 OA 的潜在治疗靶点。
