METTL14 介导的长链非编码 RNA-FAS-AS1 通过上调 ADAM8 促进骨关节炎进展。

METTL14-mediated lncRNA-FAS-AS1 promotes osteoarthritis progression by up-regulating ADAM8.

机构信息

Department of Orthopedics, The Third Affiliated Hospital of Inner Mongolia Medical University, Inner Mongolia BaoGang Hospital, Baotou, Inner Mongolia, China.

Department of Experimental Center, The Third Affiliated Hospital of Inner Mongolia Medical University, Inner Mongolia BaoGang Hospital, Baotou, Inner Mongolia, China.

出版信息

Int J Rheum Dis. 2024 Sep;27(9):e15323. doi: 10.1111/1756-185X.15323.

DOI:10.1111/1756-185X.15323

PMID:39221886

Abstract

BACKGROUND

Osteoarthritis (OA) is a prevalent degenerative disease. We explored the role and regulatory mechanisms of lncRNA-FAS-AS1 in OA progression.

METHODS

We exposed human immortalized chondrocytes to IL-1β for 24 h to induce an OA cell model. The target molecule levels were assessed using western blot and quantitative real-time PCR (RT-qPCR). Cell viability and apoptosis were measured using CCK-8 and flow cytometry. The m6A modification of FAS-AS1 was determined using MeRIP. We examined the binding relationships between FAS-AS1, Fragile X mental retardation 1 (FMR1), and A disintegrin and metalloproteinase 8 (ADAM8) using RIP and RNA pull-down. The OA animal model was established by separating the medial collateral ligament and medial meniscus. Safranin-O staining and Mankin's scale were employed to evaluate pathological changes within the cartilage.

RESULTS

FAS-AS1, METTL14, and ADAM8 were upregulated, and the JAK/STAT3 signaling pathway was activated in OA mice and IL-1β-induced chondrocytes. FAS-AS1 knockdown inhibited extracellular matrix degradation in IL-1β-induced chondrocytes; however, ADAM8 overexpression reversed this effect. FAS-AS1 maintained the stability of ADAM8 mRNA by recruiting FMR1. METTL14 knockdown repressed FAS-AS1 expression in an m6A-dependent manner. FAS-AS1 overexpression reversed the inhibitory effects of METTL14 knockdown on JAK/STAT3 signaling and cartilage damage in the OA model both in vitro and in vivo.

CONCLUSION

METTL14-mediated FAS-AS1 promotes OA progression through the FMR1/ADAM8/JAK/STAT3 axis.

摘要

背景

骨关节炎（OA）是一种常见的退行性疾病。我们探讨了长链非编码 RNA-FAS-AS1 在 OA 进展中的作用和调控机制。

方法

我们用白细胞介素-1β（IL-1β）处理人永生化软骨细胞 24 小时，以诱导 OA 细胞模型。采用 Western blot 和实时定量 PCR（RT-qPCR）检测靶分子水平。用 CCK-8 和流式细胞术检测细胞活力和凋亡。用 MeRIP 检测 FAS-AS1 的 m6A 修饰。用 RIP 和 RNA 下拉实验检测 FAS-AS1、脆性 X 智力低下蛋白 1（FMR1）和解整合素金属蛋白酶 8（ADAM8）之间的结合关系。通过分离内侧副韧带和内侧半月板建立 OA 动物模型。用番红 O 染色和 Mankin 评分评估软骨内的病理变化。

结果

在 OA 小鼠和 IL-1β诱导的软骨细胞中，FAS-AS1、METTL14 和 ADAM8 上调，JAK/STAT3 信号通路被激活。FAS-AS1 敲低抑制了 IL-1β诱导的软骨细胞中细胞外基质的降解；然而，ADAM8 的过表达逆转了这一效应。FAS-AS1 通过募集 FMR1 维持 ADAM8 mRNA 的稳定性。METTL14 敲低以 m6A 依赖性方式抑制 FAS-AS1 的表达。FAS-AS1 过表达逆转了 METTL14 敲低对 OA 模型中 JAK/STAT3 信号和软骨损伤的抑制作用，无论是在体外还是体内。