NEAT1/miR-146a-3p/TrkB/ShcB 轴调控软骨细胞的发育和功能。
NEAT1/miR-146a-3p/TrkB/ShcB axis regulates the development and function of chondrocyte.
机构信息
Department of Extremitas Superior, Luoyang Orthopedic-Traumatological Hospital Of Henan Province(Henan Provincial Orthopedic Hospital), Luoyang City, Henan Province, China.
Department of Orthopaedic Trauma and Microsurgery, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China.
出版信息
Cell Cycle. 2021 Oct;20(20):2174-2194. doi: 10.1080/15384101.2021.1974787. Epub 2021 Sep 8.
The current study aimed to explored the regulatory effect of Tropomyosin-related kinases B (TrkB) in the development and function of chondrocyte. Correlation between clinicopathological characteristics and osteoarthritis (OA) were analyzed. The expressions of TrkA, brain-derived neurotrophic factor (BDNF), TrkB, Src homolog and collagen homolog B (ShcB), and ShcC in OA cartilage tissue and IL-1β-stimulated chondrocytes from normal cartilage were determined by Western blot/qRT-PCR. After manipulating the expressions of TrkA, shTrkB, ShcB, miR-146a-3p and nuclear paraspeckle assembly transcript 1 (NEAT1), the differentiation-related molecules, and apoptosis-related molecules were examined by Western blot/qRT-PCR, and migration, invasion, proliferation, tube formation, and apoptosis rate in IL-1β-stimulated chondrocyte were examined by scratch, Transwell, colony formation, and tube formation, and flow cytometry assays, respectively. Bioinformatics, dual-luciferase and Spearman were used to analyze the binding and correlation of target genes. The findings showed that OA was related to body mass Index (BMI). The expressions of TrkA, TrkB and ShcB and NEAT1 were up-regulated in OA and IL-1β-stimulated chondrocytes, while miR-146a-3p was donwnregulated and was negatively correlated with TrkB or NEAT1. NEAT1 competed with TrkB in chondrocytes for miR-146a-3p binding. ShTrkB reversed the decrease in expressions of differentiation-related molecules, migration, invasion and proliferation, and the increase in ShcB expression and tube formation, of IL-1β-stimulated chondrocytes. Overexpressed ShcB reversed effect of shTrkB on the functions of IL-1β-stimulated chondrocytes. MiR-146a-3p inhibitor reversed effects of shTrkB on the function and apoptosis-related molecules on IL-1β-stimulated chondrocytes, while NEAT1 reversed role of miR-146a-3p. This paper demonstrated that NEAT1/miR-146a-3p/TrkB/ShcB axis regulates the development and function of chondrocyte.
本研究旨在探讨原肌球蛋白相关激酶 B (TrkB) 在软骨细胞发育和功能中的调节作用。分析了临床病理特征与骨关节炎 (OA) 的相关性。采用 Western blot/qRT-PCR 检测 OA 软骨组织和正常软骨中 IL-1β 刺激的软骨细胞中 TrkA、脑源性神经营养因子 (BDNF)、TrkB、Src 同源和胶原蛋白同源 B (ShcB) 和 ShcC 的表达。通过 Western blot/qRT-PCR 检测 TrkA、shTrkB、ShcB、miR-146a-3p 和核多泡体组装转录本 1 (NEAT1) 的表达，研究分化相关分子和凋亡相关分子的表达情况。采用划痕、Transwell、集落形成和管形成以及流式细胞术检测 IL-1β 刺激的软骨细胞的迁移、侵袭、增殖、管形成和凋亡率。采用生物信息学、双荧光素酶和 Spearman 分析靶基因的结合和相关性。结果表明，OA 与体重指数 (BMI) 有关。OA 和 IL-1β 刺激的软骨细胞中 TrkA、TrkB 和 ShcB 及 NEAT1 的表达上调，而 miR-146a-3p 下调，与 TrkB 或 NEAT1 呈负相关。NEAT1 在软骨细胞中与 TrkB 竞争 miR-146a-3p 结合。shTrkB 逆转了 IL-1β 刺激的软骨细胞中分化相关分子、迁移、侵袭和增殖的表达降低，以及 ShcB 表达和管形成的增加。过表达 ShcB 逆转了 shTrkB 对 IL-1β 刺激的软骨细胞功能的影响。miR-146a-3p 抑制剂逆转了 shTrkB 对 IL-1β 刺激的软骨细胞功能和凋亡相关分子的作用，而 NEAT1 则逆转了 miR-146a-3p 的作用。本文证明了 NEAT1/miR-146a-3p/TrkB/ShcB 轴调节软骨细胞的发育和功能。
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