敲低长链非编码 RNA HOTAIR 通过 miR-107/CXCL12 轴抑制骨关节炎软骨细胞损伤。
Knockdown of long noncoding RNA HOTAIR inhibits osteoarthritis chondrocyte injury by miR-107/CXCL12 axis.
机构信息
Department of Orthopedics, Yan'an Hospital Affiliated to Kunming Medical University, No. 245 Renmin East Road, Panlong District, Kunming, 650051, Yunnan, China.
出版信息
J Orthop Surg Res. 2021 Jun 28;16(1):410. doi: 10.1186/s13018-021-02547-7.
BACKGROUND
Osteoarthritis (OA) is a joint disease characterized via destruction of cartilage. Chondrocyte damage is associated with cartilage destruction during OA. Long noncoding RNAs (lncRNAs) are implicated in the regulation of chondrocyte damage in OA progression. This study aims to investigate the role and underlying mechanism of lncRNA homeobox antisense intergenic RNA (HOTAIR) in OA chondrocyte injury.
METHODS
Twenty-three OA patients and healthy controls without OA were recruited. Chondrocytes were isolated from OA cartilage tissues. HOTAIR, microRNA-107 (miR-107) and C-X-C motif chemokine ligand 12 (CXCL12) levels were measured by quantitative real-time polymerase chain reaction and western blot. Cell proliferation, apoptosis and extracellular matrix (ECM) degradation were measured using cell counting kit-8, flow cytometry and western blot. The target interaction was explored by bioinformatics, luciferase reporter and RNA immunoprecipitation assays.
RESULTS
HOTAIR expression was enhanced, and miR-107 level was reduced in OA cartilage samples. HOTAIR overexpression inhibited cell proliferation, but induced cell apoptosis and ECM degradation in chondrocytes. HOTAIR knockdown caused an opposite effect. MiR-107 was sponged and inhibited via HOTAIR, and knockdown of miR-107 mitigated the effect of HOTAIR silence on chondrocyte injury. CXCL12 was targeted by miR-107. CXCL12 overexpression attenuated the roles of miR-107 overexpression or HOTAIR knockdown in the proliferation, apoptosis and ECM degradation. CXCL12 expression was decreased by HOTAIR silence, and restored by knockdown of miR-107.
CONCLUSION
HOTAIR knockdown promoted chondrocyte proliferation, but inhibited cell apoptosis and ECM degradation in OA chondrocytes by regulating the miR-107/CXCL12 axis.
背景
骨关节炎（OA）是一种以软骨破坏为特征的关节疾病。软骨细胞损伤与 OA 进展过程中的软骨破坏有关。长链非编码 RNA（lncRNA）参与 OA 进展中软骨细胞损伤的调节。本研究旨在探讨 lncRNA 同源盒反义基因间 RNA（HOTAIR）在 OA 软骨细胞损伤中的作用及其潜在机制。
方法
招募 23 名 OA 患者和无 OA 的健康对照者。从 OA 软骨组织中分离软骨细胞。通过实时定量聚合酶链反应和 Western blot 检测 HOTAIR、微小 RNA-107（miR-107）和 C-X-C 基序趋化因子配体 12（CXCL12）的水平。通过细胞计数试剂盒-8、流式细胞术和 Western blot 检测细胞增殖、凋亡和细胞外基质（ECM）降解。通过生物信息学、荧光素酶报告和 RNA 免疫沉淀测定探索靶标相互作用。
结果
OA 软骨样本中 HOTAIR 表达增强，miR-107 水平降低。HOTAIR 过表达抑制软骨细胞增殖，但诱导细胞凋亡和 ECM 降解。HOTAIR 敲低则产生相反的效果。HOTAIR 通过海绵吸附并抑制 miR-107，而 miR-107 的敲低减轻了 HOTAIR 沉默对软骨细胞损伤的影响。CXCL12 是 miR-107 的靶标。CXCL12 的过表达减弱了 miR-107 过表达或 HOTAIR 敲低对增殖、凋亡和 ECM 降解的作用。HOTAIR 沉默下调 CXCL12 的表达，而 miR-107 的敲低则恢复了 CXCL12 的表达。
结论
HOTAIR 敲低通过调节 miR-107/CXCL12 轴促进 OA 软骨细胞中的软骨细胞增殖，但抑制细胞凋亡和 ECM 降解。
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