miR-653-5p 通过靶向 FGF2 并抑制 Wnt/β-catenin 通路来抑制成纤维样滑膜细胞的活力和迁移。

miR-653-5p suppresses the viability and migration of fibroblast-like synoviocytes by targeting FGF2 and inactivation of the Wnt/beta-catenin pathway.

机构信息

Department of Orthopedics, Affiliated Jianhu Hospital of Nantong University, No. 666 Nanhuan Road, Jianhu, Yancheng, 224700, Jiangsu, People's Republic of China.

出版信息

J Orthop Surg Res. 2022 Jan 4;17(1):5. doi: 10.1186/s13018-021-02887-4.

BACKGROUND

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease. Several studies reported that fibroblast-like synoviocytes (FLSs) and miRNAs are associated with RA pathogenesis. This study explored the function of miR-653-5p in the regulation of human fibroblast-like synoviocytes-rheumatoid arthritis (HFLS-RA) cells.

METHODS

The mRNA and protein levels of genes were measured by RT-qPCR and western blot, respectively. MTT, wound healing, and invasion assays were used to evaluate the viability and metastasis of FLSs. Luciferase reporter and RNA pull-down assays were employed to determine the interaction between miR-653-5p and FGF2.

RESULTS

RT-qPCR results demonstrated that miR-653-5p expression was decreased and FGF2 level was increased in synovial tissues and FLSs of RA. Moreover, the viability and metastasis of FLSs were accelerated by miR-653-5p addition, which was restrained by miR-653-5p suppression. Furthermore, we demonstrated that levels of Rac1, Cdc42, and RhoA were decreased after miR-653-5p addition. Besides, luciferase reporter and RNA pull-down assays implied that miR-653-5p targeted the 3'-UTR of FGF2. Functional assays showed that FGF2 overexpression neutralized the suppressive effects of miR-653-5p addition on HFLS-RA cell viability, metastasis, and the levels of Rho family proteins. Meanwhile, the levels of β-catenin, cyclin D1, and c-myc were declined by miR-653-5p supplementation, but enhanced by FGF2 addition.

CONCLUSION

In sum, we manifested that miR-653-5p restrained HFLS-RA cell viability and metastasis via targeting FGF2 and repressing the Wnt/beta-Catenin pathway.

背景

类风湿关节炎（RA）是一种慢性系统性自身免疫性疾病。几项研究报道成纤维细胞样滑膜细胞（FLS）和 microRNA（miRNA）与 RA 的发病机制有关。本研究探讨了 miR-653-5p 在调节人成纤维细胞样滑膜细胞-类风湿关节炎（HFLS-RA）细胞中的功能。

方法

通过 RT-qPCR 和 Western blot 分别测量基因的 mRNA 和蛋白水平。MTT、划痕愈合和侵袭实验用于评估 FLS 的活力和转移。荧光素酶报告和 RNA 下拉实验用于确定 miR-653-5p 和 FGF2 之间的相互作用。

结果

RT-qPCR 结果表明，miR-653-5p 在滑膜组织和 RA 的 FLS 中表达降低，FGF2 水平升高。此外，miR-653-5p 的添加加速了 FLS 的活力和转移，而 miR-653-5p 的抑制则抑制了其活力和转移。此外，我们证明 miR-653-5p 加药后 Rac1、Cdc42 和 RhoA 的水平降低。此外，荧光素酶报告和 RNA 下拉实验表明，miR-653-5p 靶向 FGF2 的 3'-UTR。功能测定表明，FGF2 的过表达中和了 miR-653-5p 加药对 HFLS-RA 细胞活力、转移和 Rho 家族蛋白水平的抑制作用。同时，miR-653-5p 的补充降低了 β-连环蛋白、细胞周期蛋白 D1 和 c-myc 的水平，但增强了 FGF2 的添加。