Department of Orthopedics, The First Affiliated Hospital of Soochow University, Suzhou, China.
Department of Orthopedics, The Third Affiliated Hospital of Shanghai University; The Wenzhou Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou, China.
J Orthop Surg (Hong Kong). 2024 May-Aug;32(2):10225536241254588. doi: 10.1177/10225536241254588.
The abnormal function and survival of chondrocytes result in articular cartilage failure, which may accelerate the onset and development of osteoarthritis (OA). This study is aimed to investigate the role of LINC01094 in chondrocyte apoptosis.
The viability and apoptosis of lipopolysaccharide (LPS)-induced chondrocytes were evaluated through CCK-8 assay and flow cytometry analysis, respectively. The expression levels of LINC01094, miR-577 and MTF1 were detected by qRT-PCR. Dual luciferase reporter tests were implemented for the verification of targeted relationships among them. Western blotting was employed to measure the levels of pro-apoptotic proteins (Caspase3 and Caspase9).
The viability of LPS-induced chondrocytes was overtly promoted by loss of LINC01094 or miR-577 upregulation, but could be repressed via MTF1 overexpression. The opposite results were observed in apoptosis rate and the levels of Caspase3 and Caspase9. LINC01094 directly bound to miR-577, while MTF1 was verified to be modulated by miR-577. Both LINC01094 and MTF1 were at high levels, whereas miR-577 was at low level in OA synovial fluid and LPS-induced chondrocytes. Furthermore, the highly expressed miR-577 abolished the influences of MTF1 overexpression on LPS-induced chondrocytes.
Silencing of LINC01094 represses the apoptosis of chondrocytes through upregulating miR-577 expression and downregulating MTF1 levels, providing a preliminary insight for the treatment of OA in the future.
软骨细胞的异常功能和存活导致关节软骨失效，这可能加速骨关节炎（OA）的发生和发展。本研究旨在探讨 LINC01094 在软骨细胞凋亡中的作用。
通过 CCK-8 检测和流式细胞术分析分别评估脂多糖（LPS）诱导的软骨细胞的活力和凋亡。通过 qRT-PCR 检测 LINC01094、miR-577 和 MTF1 的表达水平。通过双荧光素酶报告实验验证它们之间的靶向关系。Western blot 用于测量促凋亡蛋白（Caspase3 和 Caspase9）的水平。
LINC01094 缺失或 miR-577 上调明显促进 LPS 诱导的软骨细胞活力，但通过 MTF1 过表达可抑制其活力。凋亡率和 Caspase3 和 Caspase9 的水平则观察到相反的结果。LINC01094 直接与 miR-577 结合，而 MTF1 被证实受 miR-577 调控。OA 滑膜液和 LPS 诱导的软骨细胞中 LINC01094 和 MTF1 水平较高，而 miR-577 水平较低。此外，高表达的 miR-577 消除了 MTF1 过表达对 LPS 诱导的软骨细胞的影响。
沉默 LINC01094 通过上调 miR-577 表达和下调 MTF1 水平抑制软骨细胞凋亡，为未来 OA 的治疗提供了初步见解。
