lncRNA MEG3作为ceRNA调控miR-16在骨性关节炎中的作用及机制研究

Long non-coding RNAs (lncRNA), which may act as competing endogenous RNAs (ceRNAs), play a critical role in the pathological process of many diseases. However, their biological role and underlying molecular mechanism in osteoarthritis (OA) remains largely undefined. Our previous study has found that lncRNA MEG3 down-regulation is inversely correlated with the upregulation of miR-16 in OA, which were also predicted by bioinfomatics. We also found that MEG3 could improve the capacity of anabolism in chondrocytes and supress the expression of miR-16. Furthermore, the result of bioinfomatics and luciferase reporter assay suggests that MEG3 may function as a ceRNA to regulate the expression of BMP/TGFβ/Smad pathways (SMAD3 and SOX5,especially) by sponging miR-16. Hereby, we propose the hypothesis: MEG3 functions as a ceRNA to regulate BMP/TGFβ/Smad pathways by sponging miR-16 and affects the progression of OA. To investigate the effect and mechanism of MEG3 in the regulation of miR-16, BMP/TGFβ/Smad pathways and the progression of OA, we utilize a series of experiments in vitro and in vivo, including examining clinical tissue samples, constructing MEG3 overexpression/deficiency chondrocyte model, establishment of the mice OA model, RNA Binding Protein Immunoprecipitation (RIP) and luciferase reporter assays. This study has important significance to clarify the pathogenisis of OA and will provide us new ideas and theoretical basis to early diagnosis and targeted therapy in OA.

lncRNA可以作为竞争性内源RNA（ceRNA）参与疾病的病理进程，而其在骨性关节炎（OA）及软骨细胞中的作用尚未阐明。我们前期发现lncRNA MEG3在OA软骨中低表达且与miR-16的表达负相关；体外实验发现MEG3可以抑制miR-16并促进软骨细胞合成代谢；生物信息学和荧光报告基因实验表明MEG3可能吸附miR-16并作用于TGFβ/BMP/Smad信号通路中SMAD3 和SOX5。据此我们提出假设：软骨细胞中MEG3作为ceRNA吸附miR-16调控TGFβ/BMP/Smad信号通路中SMAD3、SOX5的表达，最终影响OA的发生与进展。本研究拟通过收集临床标本，构建MEG3过/缺失表达细胞模型及小鼠OA模型，采用RNA免疫共沉淀、荧光报告基因等技术，多层面、系统的研究OA中MEG3的生物作用及其作为ceRNA的分子调控机制，以期为OA早期诊断和靶向治疗提供新的思路和实验基础。

lncRNA可以作为竞争性内源RNA（ceRNA）参与疾病的病理进程，而其在骨性关节炎（OA）及软骨细胞中的作用尚未阐明。该项目属于申请人前期研究成果的延续和深入，探索MEG3对骨关节炎的影响，以及通过吸附miR-16-5p调控下游基因SMAD3的功能作用及相关机制。证实MEG3-miR-16-5p-SMAD3在骨关节炎发生发展中的机制。本研究进一步将生物信息学的技术运用于骨科，首先通过数据挖掘找到了骨肉瘤的复发转移相关分子分型（PQBP1高表达、 PCK2低表达）。并利用相关的研究方法找到了骨关节炎中两个新的关键因子lncRNA CTD-2574D22.4与PMS2L2。发现抑制lncRNA CTD-2574D22.4可减轻LPS诱导的软骨细胞凋亡和炎症损伤；而lncRNA PMS2L2通过激活软骨细胞自噬抑制骨关节炎进展。目前发表4篇相关SCI论文，并仍有一篇在投稿。研究结果对了解OA的发生发展机理及寻找OA的可能治疗靶点具有重要的意义。