Linc-ROR 通过调节 SOX9 的表达促进间充质干细胞的软骨生成和软骨形成。

Linc-ROR promotes mesenchymal stem cells chondrogenesis and cartilage formation via regulating SOX9 expression.

机构信息

Department of Orthopaedics & Traumatology, Stem Cells and Regenerative Medicine Laboratory, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, SAR, PR China; MOE Key Laboratory for Regenerative Medicine, School of Biomedical Sciences, The Chinese University of Hong Kong, SAR, PR China; Department of Orthopaedics and Traumatology, People's Hospital of Baoan District, Shenzhen, PR China.

出版信息

Osteoarthritis Cartilage. 2021 Apr;29(4):568-578. doi: 10.1016/j.joca.2020.12.020. Epub 2021 Jan 22.

DOI:10.1016/j.joca.2020.12.020

PMID:33485931

Abstract

OBJECTIVE

The present study is to characterize the role of long intergenic non-coding RNA, regulator of reprogramming (linc-ROR) in bone marrow mesenchymal stem cell (BMSCs) chondrogenesis, cartilage formation and OA development.

METHODS

Linc-ROR expression pattern in articular cartilage tissue sample from OA patients were studied by real-time PCR. Linc-ROR lentivirus mediated BMSCs were constructed. In vitro micromass cultured BMSCs chondrogenesis or in vivo MeHA hydrogel encapsulated BMSCs cartilage formation activity were studied. Linc-ROR associating miRNAs which repressed SOX9 expression were characterized by luciferase assay, real-time PCR and Western blot. Linc-ROR was co-transfected with miRNAs into BMSCs to study its rescue effect on SOX9 expression and chondrogenesis activity.

RESULTS

Linc-ROR was down-regulated in articular cartilage tissue from OA patients and was positively correlated with the expression level of SOX9 (R = 0.43). Linc-ROR expression was upregulated during BMSCs chondrogenesis. Linc-ROR ectopic expression significantly promoted in vitro BMSCs chondrogenesis and in vivo cartilage formation activities as revealed by safranin O, alcian blue and COL II staining. The mRNA expression level of chondrogenesis markers including COL II, SOX9 and ACAN were increased, and the hypertrophy markers MMP13 and COL X were decreased upon linc-ROR overexpression in BMSCs. Linc-ROR functioned as a miRNA sponge for miR-138 and miR-145. Both miR-138 and miR-145 suppressed BMSCs chondrogenesis activity and SOX9 expression, while co-expression of linc-ROR displayed a rescuing effect.

CONCLUSIONS

Taken together, linc-ROR modulated BMSCs chondrogenesis differentiation and cartilage formation by acting as a competing endogenous RNA for miR-138 and miR-145 and activating SOX9 expression. Linc-ROR could be considered as a new diagnostic and therapeutic target for OA treatment.

摘要

目的

本研究旨在探讨长链非编码 RNA，重编程调节因子（linc-ROR）在骨髓间充质干细胞（BMSCs）软骨分化、软骨形成和 OA 发展中的作用。

方法

通过实时 PCR 研究 OA 患者关节软骨组织样本中 linc-ROR 的表达模式。构建 linc-ROR 慢病毒转染的 BMSCs。研究体外微团培养 BMSCs 软骨分化或体内 MeHA 水凝胶包埋 BMSCs 软骨形成活性。通过荧光素酶报告基因、实时 PCR 和 Western blot 分析抑制 SOX9 表达的 linc-ROR 相关 miRNA。将 linc-ROR 与 miRNA 共转染入 BMSCs 中，研究其对 SOX9 表达和软骨分化活性的挽救作用。

结果

linc-ROR 在 OA 患者关节软骨组织中表达下调，与 SOX9 的表达水平呈正相关（R=0.43）。BMSCs 软骨分化过程中 linc-ROR 表达上调。linc-ROR 异位表达显著促进体外 BMSCs 软骨分化和体内软骨形成活性，经 safranin O、阿利新蓝和 COL II 染色证实。BMSCs 中 linc-ROR 过表达后，软骨分化标志物 COL II、SOX9 和 ACAN 的 mRNA 表达水平升高，肥大标志物 MMP13 和 COL X 表达水平降低。linc-ROR 作为 miR-138 和 miR-145 的 miRNA 海绵发挥作用。miR-138 和 miR-145 均抑制 BMSCs 软骨分化活性和 SOX9 表达，而 linc-ROR 的共表达显示出挽救作用。