1Department of Rheumatology and Immunology, Cangzhou Central Hospital, Cangzhou, 061000 People's Republic of China.
2The Second Nephrology Department, Cangzhou Central Hospital, Cangzhou, 061000 People's Republic of China.
Cell Mol Biol Lett. 2019 Apr 19;24:8. doi: 10.1186/s11658-018-0130-0. eCollection 2019.
This study aimed to investigate the effects of miR-613 on the proliferation, invasion and apoptosis of rheumatoid arthritis synovial fibroblasts (RASFs).
Synovial tissue samples were collected from 20 rheumatoid arthritis (RA) patients and 10 patients with joint trauma undergoing joint replacement surgery. The RASFs were isolated and cultured. MiR-613 and DKK1 expression in both synovial tissues and cells was detected using quantitative real-time PCR (qRT-PCR). Dual luciferase reporter gene assay was employed to evaluate the effect of miR-613 on the luciferase activity of DKK1. Then RASFs were transfected with miR-613 mimics, si-DKK1 and pcDNA-DKK1. Changes in cellular proliferation, invasion and apoptosis were detected through BrdU assay, Transwell invasion assay and flow cytometry analysis, respectively.
MiR-613 was significantly down-regulated in RA tissues and RASFs compared to normal tissues and cells, whereas DKK1 was up-regulated in RA tissues and RASFs. Dual luciferase reporter gene assay showed that miR-613 could specifically bind to the 3'UTR of DKK1 and significantly inhibit the luciferase activity. Moreover, miR-613 significantly reduced the expression of DKK1. Overexpression of miR-613 or knockdown of DKK1 suppressed proliferation and invasion of RASFs, and induced RASF apoptosis. The reverse results were observed when DKK1 was up-regulated in miR-613-overexpressing RASFs.
MiR-613 can inhibit proliferation and invasion and induce apoptosis of RASFs by directly targeting DKK1 expression.
本研究旨在探讨 miR-613 对类风湿关节炎滑膜成纤维细胞（RASFs）增殖、侵袭和凋亡的影响。
收集 20 例类风湿关节炎（RA）患者和 10 例关节置换术的关节创伤患者的滑膜组织标本。分离培养 RASFs。采用实时定量 PCR（qRT-PCR）检测滑膜组织和细胞中 miR-613 和 DKK1 的表达。双荧光素酶报告基因实验评估 miR-613 对 DKK1 荧光素酶活性的影响。然后用 miR-613 模拟物、si-DKK1 和 pcDNA-DKK1 转染 RASFs。通过 BrdU 检测、Transwell 侵袭实验和流式细胞术分析分别检测细胞增殖、侵袭和凋亡的变化。
与正常组织和细胞相比，miR-613 在 RA 组织和 RASFs 中显著下调，而 DKK1 在 RA 组织和 RASFs 中上调。双荧光素酶报告基因实验显示，miR-613 可特异性结合 DKK1 的 3'UTR，并显著抑制荧光素酶活性。此外，miR-613 显著降低了 DKK1 的表达。过表达 miR-613 或敲低 DKK1 抑制 RASFs 的增殖和侵袭，并诱导 RASF 凋亡。在 miR-613 过表达的 RASFs 中上调 DKK1 则得到相反的结果。
miR-613 可通过直接靶向 DKK1 的表达抑制 RASFs 的增殖、侵袭并诱导其凋亡。
