Effects of miR-214 on the proliferation and apoptosis of rheumatoid arthritis synovial fibroblasts by targeting sex determining region Y-box 4
10.3760/cma.j.c141217-20201209-00471
- VernacularTitle:微RNA-214靶向性别决定区Y-box 4对类风湿关节炎滑膜成纤维细胞增殖凋亡的影响
- Author:
Xiaoying WEI
1
;
Juntao WANG
;
Jingxia TIAN
;
Xinping WEI
Author Information
1. 河南省商丘市第一人民医院肾病风湿科 476100
- Keywords:
Arthritis, rheumatoid;
MicroRNAs;
Sex-determining region Y-protein;
Nnuclear transcription factor-κB signaling pathway;
Proliferation;
Apoptosis
- From:
Chinese Journal of Rheumatology
2021;25(7):455-460,c7-1
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of miR-214 targeting sex determining region Y-box 4 (SOX4) on the proliferation and apoptosis of rheumatoid arthritis (RA) synovial fibroblasts.Methods:Synovial tissue samples from 45 patients with rheumatoid arthritis and 30 patients with joint trauma who underwent joint replacement surgery from 2017 to 2019 in Shangqiu First People's Hospital samples were collected. The expression levels of miR-214 and SOX4 in synovial tissues were detected by real time quantitative polymerase chain reaction (RT-qPCR); The expression levels of serum rheumatoid factor (RF) and C-reactive protein (CRP) were detected by enzyme-linked immunosorbent assay (ELISA) test. The correlation between miR-214 or SOX4 and RF, CRP, erythrocyte sedimentation rate (ESR) was tested by Pearson's analysis. Human fibroblast-like synoviocytes (HFLS)-RA cells were derived from the control group, miR-214 mimic group, mimic-NC group and miR-214 mimic+pcDNA3.1-SOX4 group and se-parated by Lipofectamine 3 000. The cell viability of each group was detected by CCK8, the apoptosis of each group was detected by flow cytometry, the target relationship was verified by dual luciferase experiment, Western-blot was used to detect the protein expression levels of related indicators. Independent sample t test was used for the comparison between the two groups, one-way nalysis of variance (ANOVA) was used for multi-group comparison, and LSD- t test was used for the comparison between the two groups. Results:Compared with the control group (4.6±0.7, 1.9±0.7) and HFLS cells (1.00±0.06, 1.00±0.09), miR-214 was expressed lower in RA tissues (2.6±0.9) and HFLS-RA (0.30±0.05) ( t=10.026, P<0.05; t=15.815, P<0.05). Compared with SOX4 was highly expressed in RA tissues (4.6±0.9) and HFLS-RA cells (3.89±0.41) ( t=14.772, P<0.05; t=12.020, P<0.05). Serum RF, ESR and CRP expression levels in RA group [(46±7) U/ml, (46.4±9.6) mm/1 h, (34.8±9.8) mg/ml] were signifi-cantly higherthan those in the control group [(16±4) U/ml, (9.2±2.0) mm/1 h, (2.1±0.7) mg/ml] ( t=22.906, P<0.05; t=25.338, P<0.05; t=22.314, P<0.05). miR-214 was negatively correlated with serum RF, CRP and ESR was negatively correlated with miRNA( r=-0.574, P<0.05; r=-0.448, P<0.05; r=-0.549, P<0.05), while was positively correlated with SOX4 ( r=0.492, P<0.05; r=0.369, P<0.05; r=0.325, P<0.05). Compared with the control group and the mimic-NC group, cell viability, Ki-67 and p-p65 protein expression decreased significantly after miR-214 mimic transfection for 24 h ( F24 h=16.980, P<0.05;, F48 h=42.735, P<0.05; F72 h=164.448, P<0.05; FKi-67=290.112, P<0.05; Fp-p65/p65=223.548, P<0.05), while the apoptosis rate and Bax protein expression increased significantly ( F=344.360, P<0.05; F=106.376, P<0.05). Overexpression of SOX4, reverses the effect of miR-214. Conclusion:miR-214 targets SOX4 to inhibit the proliferation of synovial fibroblasts and promote apoptosis in rheumatoid arthritis. This may relate to the inhibition of NF-κB signaling pathway by miR-214.
