Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, Hubei Province, China.
Cartilage. 2021 Dec;13(2_suppl):1412S-1420S. doi: 10.1177/19476035211003324. Epub 2021 Apr 9.
Osteoarthritis (OA) is a common joint disorder, accompanied by extracellular matrix (ECM) degradation. Reportedly, long noncoding RNAs (lncRNAs) are involved in OA pathogenesis. However, the role of lncRNA FYVE, RhoGEF, and PH domain containing 5 antisense RNA 1 (FGD5-AS1) in OA development is still not fully clarified. This study was aimed to clarify the role of FGD5-AS1 in OA.
FGD5-AS1 and miR-302d-3p expression levels were determined in cartilage tissues and chondrocytes by quantitative real-time polymerase chain reaction (qRT-PCR). Chondrocytes (C20/A4 cells) were stimulated with interleukin 1β (IL-1β) to mimic the inflammatory environment of OA. Cell viability was detected by cell counting kit-8 and 5-ethynyl-2'-deoxyuridine assays. Cell apoptosis was measured by the caspase-3 activity assay and flow cytometry. Transforming growth factor beta receptors II (TGFBR2), matrix metalloproteinase 13 (MMP-13), and ADAM metallopeptidase with thrombospondin type 1 motif 5 expression levels were examined by qRT-PCR or Western blot. The regulatory relationships among FGD5-AS1, miR-302d-3p, and TGFBR2 were predicted by the StarBase v2.0, miRanda, miRDB, and TargetScan databases, and confirmed by dual-luciferase reporter assay and RNA immunoprecipitation assay.
FGD5-AS1 and TGFBR2 expression levels were downregulated while miR-302d-3p expression was increased in cartilage tissues of OA patients. Knocking down FGD5-AS1 inhibited the viability of C20/A4 cells but induced apoptosis and ECM degradation, while FGD5-AS1 overexpression exerted opposite effects. MiR-302d-3p was identified as a target of FGD5-AS1, and TGFBR2 was identified as a target of miR-302d-3p. FGD5-AS1 positively regulated TGFBR2 expression by repressing miR-302d-3p expression, and miR-302d-3p inhibition or TGFBR2 restoration reversed the changes of cell viability, apoptosis, and ECM degradation induced by FGD5-AS1 knockdown.
FGD5-AS1 can probably inhibit OA progression by regulating miR-302d-3p/TGFBR2 axis.
骨关节炎（OA）是一种常见的关节疾病，伴随着细胞外基质（ECM）降解。据报道，长链非编码 RNA（lncRNA）参与 OA 的发病机制。然而，lncRNA FYVE、RhoGEF 和 PH 结构域包含 5 个反义 RNA 1（FGD5-AS1）在 OA 发展中的作用仍未完全阐明。本研究旨在阐明 FGD5-AS1 在 OA 中的作用。
通过实时定量聚合酶链反应（qRT-PCR）检测软骨组织和软骨细胞中的 FGD5-AS1 和 miR-302d-3p 表达水平。用白细胞介素 1β（IL-1β）刺激软骨细胞（C20/A4 细胞）模拟 OA 的炎症环境。通过细胞计数试剂盒-8 和 5-乙炔基-2'-脱氧尿苷测定法检测细胞活力。通过 caspase-3 活性测定法和流式细胞术测量细胞凋亡。通过 qRT-PCR 或 Western blot 检测转化生长因子β受体 II（TGFBR2）、基质金属蛋白酶 13（MMP-13）和 ADAM 金属肽酶与血小板反应蛋白 1 型基序 5（ADAMTS5）的表达水平。FGD5-AS1、miR-302d-3p 和 TGFBR2 之间的调控关系通过 StarBase v2.0、miRanda、miRDB 和 TargetScan 数据库预测，并通过双荧光素酶报告基因检测和 RNA 免疫沉淀检测证实。
OA 患者软骨组织中 FGD5-AS1 和 TGFBR2 表达下调，而 miR-302d-3p 表达上调。敲低 FGD5-AS1 抑制 C20/A4 细胞的活力，但诱导细胞凋亡和 ECM 降解，而 FGD5-AS1 过表达则产生相反的效果。miR-302d-3p 被鉴定为 FGD5-AS1 的靶标，TGFBR2 被鉴定为 miR-302d-3p 的靶标。FGD5-AS1 通过抑制 miR-302d-3p 表达来正向调节 TGFBR2 表达，而 FGD5-AS1 抑制或 TGFBR2 恢复可逆转 FGD5-AS1 敲低引起的细胞活力、凋亡和 ECM 降解变化。
FGD5-AS1 可能通过调节 miR-302d-3p/TGFBR2 轴抑制 OA 进展。
