长链非编码 RNA PVT1 的敲低通过调节 miR-543 依赖的 SCUBE2 诱导类风湿关节炎成纤维样滑膜细胞凋亡。

Knockdown of long non-coding RNA PVT1 induces apoptosis of fibroblast-like synoviocytes through modulating miR-543-dependent SCUBE2 in rheumatoid arthritis.

机构信息

Department of Rheumatism, Linyi Central Hospital, No. 17, Jiankang Road, Yishui Town, Linyi, 276400, Shandong, People's Republic of China.

出版信息

J Orthop Surg Res. 2020 Apr 15;15(1):142. doi: 10.1186/s13018-020-01641-6.

BACKGROUND

Rheumatoid arthritis (RA), a kind of autoimmune disorder, is featured by many physical symptoms and proliferation of fibroblast-like synoviocytes (FLSs). The relevance of long non-coding RNAs (lncRNAs) in the progression of RA has been probed. Hence, the goal of this report was to investigate the action of plasmacytoma variant translocation 1 (PVT1), a lncRNA, in FLSs and the basic mechanism.

METHODS

Initially, RA rats were developed to evaluate the expression of PVT1, microRNA-543 (miR-543), and signal peptide-CUB-EGF-like containing protein 2 (SCUBE2) in synovial tissues. Enhancement or loss of PVT1 or miR-543 was achieved to explore their effects on proliferation, cell cycle, and apoptosis of FLSs. The interaction between PVT1 and miR-543 and between miR-543 and its putative target SCUBE2 was examined to elucidate the correlations. Finally, the protein expression of proliferation- and apoptosis-associated genes were assessed by western blot assays.

RESULTS

PVT1 was overexpressed in synovial tissues from RA patients through microarray expression profiles. The PVT1 and SCUBE2 expression was boosted, and miR-543 was reduced in synovial tissues of rats with RA. PVT1 specifically bound to miR-543, and miR-543 negatively regulated SCUBE2 expression. Overexpression of PVT1 or silencing of miR-543 enhanced SCUBE2 expression, thereby promoting proliferation and interleukin-1β (IL-1β) secretion, while inhibiting apoptosis rate of FLSs. Conversely, si-SCUBE2 reversed the role of miR-543 inhibitor.

CONCLUSION

The key findings support that PVT1 knockdown has the potency to hinder RA progression by inhibiting SCUBE2 expression to sponge miR-543.

背景

类风湿关节炎（RA）是一种自身免疫性疾病，其特征是多种身体症状和成纤维样滑膜细胞（FLS）的增殖。长链非编码 RNA（lncRNA）在 RA 进展中的相关性已被探讨。因此，本报告的目的是研究浆细胞瘤变异易位 1（PVT1）一种 lncRNA 在 FLS 中的作用及其基本机制。

方法

首先，建立 RA 大鼠模型，以评估滑膜组织中 PVT1、微小 RNA-543（miR-543）和信号肽-CUB-EGF 样结构域蛋白 2（SCUBE2）的表达。增强或缺失 PVT1 或 miR-543，以探讨它们对 FLS 增殖、细胞周期和凋亡的影响。检测 PVT1 与 miR-543 之间以及 miR-543 与其假定靶标 SCUBE2 之间的相互作用，以阐明相关性。最后，通过 Western blot 检测增殖和凋亡相关基因的蛋白表达。

结果

通过微阵列表达谱，在 RA 患者的滑膜组织中发现 PVT1 过表达。在 RA 大鼠的滑膜组织中，PVT1 和 SCUBE2 的表达增加，而 miR-543 的表达减少。PVT1 特异性结合 miR-543，miR-543 负调控 SCUBE2 的表达。过表达 PVT1 或沉默 miR-543 增强了 SCUBE2 的表达，从而促进了 FLSs 的增殖和白细胞介素-1β（IL-1β）的分泌，同时抑制了 FLSs 的凋亡率。相反，si-SCUBE2 逆转了 miR-543 抑制剂的作用。