Department of Orthopedics, Lihuili Hospital Affiliated to Ningbo University, Ningbo, Zhejiang 315211, P.R. China.
Department of Orthopedics, Hwa Mei Hospital, University of Chinese Academy of Sciences, Ningbo, Zhejiang 315100, P.R. China.
Mol Med Rep. 2022 Jan;25(1). doi: 10.3892/mmr.2021.12536. Epub 2021 Nov 19.
Numerous studies have previously demonstrated that long non‑coding RNAs (lncRNAs) serve an important regulatory role in osteoarthritis (OA). In particular, the lncRNA family with sequence similarity 201 member A (FAM201A) was previously found to be downregulated in necrotic femoral head samples. However, the role of FAM201A in IL‑1β‑induced chondrocyte injury remains unclear. It was hypothesized that FAM201A may exert a protective effect on IL‑1β‑induced chondrocyte injury in OA by sponging microRNAs (miRNAs/miRs). The purpose of the present study was to explore the role and molecular mechanism of FAM201A in IL‑1β‑induced chondrocyte injury. A model of OA was established by stimulation C‑28/I2 cell with IL‑1β . The expression levels of FAM201A following IL‑1β‑induced chondrocyte injury were detected via reverse transcription‑quantitative PCR. Luciferase reporter assay was used to assess the possible associations among FAM201A, miR‑146a‑5p and POU class 2 homeobox 1 (POU2F1). Chromatin immunoprecipitation assay was performed to analyze the interaction between POU2F1 and miR‑146a‑5p. ELISA, TUNEL and western blotting were performed to measure the level of inflammation, lactate dehydrogenase release, apoptosis and the expression of apoptosis‑related proteins (Bcl‑2, Bax, cleaved caspase 3 and cleaved caspase 9), respectively. The expression levels of FAM201A were found to be downregulated following IL‑1β‑induced chondrocyte injury. Overexpression of FAM201A exerted a protective effect against IL‑1β‑induced chondrocyte injury. In addition, FAM201A could upregulate the expression levels of POU2F1 by sponging miR‑146a‑5p. Further experiments revealed that POU2F1 could bind to the promoter region of FAM201A and subsequently regulate the expression levels of POU2F1, indicating a role for the FAM201A/miR‑146a‑5p/POU2F1 positive feedback loop in IL‑1β‑induced chondrocyte injury. The present study revealed the protective effects of the FAM201A/miR‑146a‑5p/POU2F1 positive feedback loop on IL‑1β‑induced chondrocyte injury and provided a potential therapeutic target for OA.
先前的大量研究表明，长链非编码 RNA（lncRNA）在骨关节炎（OA）中发挥重要的调控作用。特别是，先前发现具有序列相似性 201 成员 A（FAM201A）的 lncRNA 家族在坏死股骨头样本中下调。然而，FAM201A 在 IL-1β诱导的软骨细胞损伤中的作用尚不清楚。研究假设 FAM201A 可能通过海绵吸附 microRNAs（miRNAs/miRs）对 IL-1β诱导的软骨细胞损伤发挥保护作用。本研究旨在探讨 FAM201A 在 IL-1β诱导的软骨细胞损伤中的作用及分子机制。通过用 IL-1β刺激 C-28/I2 细胞建立 OA 模型。通过逆转录-定量 PCR 检测 IL-1β诱导的软骨细胞损伤后 FAM201A 的表达水平。利用荧光素酶报告基因实验评估 FAM201A、miR-146a-5p 和 POU 类 2 同源框 1（POU2F1）之间的可能关联。进行染色质免疫沉淀实验分析 POU2F1 与 miR-146a-5p 的相互作用。通过 ELISA、TUNEL 和 Western blot 分别测定炎症水平、乳酸脱氢酶释放、凋亡以及凋亡相关蛋白（Bcl-2、Bax、cleaved caspase 3 和 cleaved caspase 9）的表达水平。结果发现，IL-1β 诱导的软骨细胞损伤后 FAM201A 的表达水平下调。FAM201A 的过表达对 IL-1β 诱导的软骨细胞损伤具有保护作用。此外，FAM201A 可通过海绵吸附 miR-146a-5p 而上调 POU2F1 的表达水平。进一步的实验表明，POU2F1 可以结合 FAM201A 的启动子区域，进而调节 POU2F1 的表达水平，表明 FAM201A/miR-146a-5p/POU2F1 正反馈环在 IL-1β 诱导的软骨细胞损伤中发挥作用。本研究揭示了 FAM201A/miR-146a-5p/POU2F1 正反馈环对 IL-1β 诱导的软骨细胞损伤的保护作用，并为 OA 提供了一个潜在的治疗靶点。
