SOX9 promotes proliferation of fibroblast-like synoviocytes in rheumatoid arthritis by inhibiting miR-204/-211

doi:10.16138/j.1673-6087.2025.06.04

Abstract

Abstract:

Objective To investigate the effect of sex determining region Y-box transcription factor 9 (SOX9) on the function of fibroblast-like synoviocyte (FLS) in rheumatoid arthritis (RA) by inhibiting micro-ribonucleic acid (miRNA)-204 and -211, and to explore the underlying mechanism. Methods A collagene-induced arthritis (CIA) model was established by type Ⅱ collagen induction. FLS were isolated from the knee joint synovial tissue of CIA mice by digestion with type Ⅳ collagenase. The expression level of Sox9 in FLS was detected by quantitative real-time polymerase chain reaction (qPCR). Small interfering ribonucleic acid (siRNA) was used to inhibit Sox9 expression, and cell proliferation was assessed by CCK-8 assay and 5-ethynyl-2’-deoxyuridine (EdU) staining. Cell senescence was analyzed by senescence-associated β-galactosidase (SA-β-gal) staining, and the expression of cyclin-dependent kinase inhibitor (CKI) p16, p21, p53 and miR-204, miR-211 was detected by qPCR. The HDOCK online server (http://hdock.phys.hust.edu.cn/) was used to analyze the binding potential between miR-204/miR-211 and SOX9. Results Compared with FLS from normal mice, the messenger ribonucleic acid (mRNA) level of SOX9 was significantly increased in CIA-FLS. Knock down of Sox9 can promote cell senescence, thereby inhibiting FLS proliferation. Both miR-204 and miR-211 could stably bind to SOX9. Knockdown of Sox9 in CIA-FLS can increase the expression of miR-204/-211. Conclusions SOX9 can promote the proliferation of CIA-FLS and inhibit their senescence by down-regulating the expression of miR-204 and miR-211.

Key words: SOX9, miR-204/-211, Fibroblast-like synoviocyte

CLC Number:

R593.22

YANG Yilei, LIU Jing, FAN Kaijian, WANG Tingyu. SOX9 promotes proliferation of fibroblast-like synoviocytes in rheumatoid arthritis by inhibiting miR-204/-211[J]. Journal of Internal Medicine Concepts & Practice, 2026, 20(06): 449-456.

Figures/Tables 5

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References 25

[1]	Giles JT. Extra-articular manifestations and comorbidity in rheumatoid arthritis: potential impact of pre-rheumatoid arthritis prevention[J]. Clin Ther, 2019, 41(7):1246-1255. doi: 10.1016/j.clinthera.2019.04.018
[2]	Aletaha D, Smolen JS. Diagnosis and management of rheumatoid arthritis: a review[J]. JAMA, 2018, 320(13):1360-1372. doi: 10.1001/jama.2018.13103
[3]	Masoumi M, Bashiri H, Khorramdelazad H, et al. Destructive roles of fibroblast-like synoviocytes in chronic inflammation and joint damage in rheumatoid arthritis[J]. Inflammation, 2021, 44(2):466-479. doi: 10.1007/s10753-020-01371-1
[4]	Yoshitomi H. Regulation of immune responses and chronic inflammation by fibroblast-like synoviocytes[J]. Front Immunol, 2019, 10: 1395. doi: 10.3389/fimmu.2019.01395
[5]	陈伟达, 石玮, 杨继国, 等. 川续断总皂苷调节miR-19a对大鼠骨关节炎软骨细胞SOX9/NF-κB信号通路的影响[J]. 中国药师, 2020, 23(5):824-829. doi: 10.3969/j.issn.1008-049X.2020.05.007
	Chen WD, Shi W, Yang JG, et al. Effects of total saponins of dipsacus asperoides on SOX9/NF-κB signaling pathway in rat's osteoarthritic chondrocytes through regulating miR-19a[J]. China Pharmacist, 2020, 23(5):824-829. doi: 10.3969/j.issn.1008-049X.2020.05.007
[6]	卓群豪, 张伟娜, 李舰, 等. 膝关节腔内注射Sox9转染骨髓间充质干细胞修复膝关节骨关节炎[J]. 中国组织工程研究, 2017, 21(5):736-741. doi: 10.3969/j.issn.2095-4344.2017.05.014
	Zhuo QH, Zhang WN, Li J, et al. Intra-articular injection of Sox9-transfected bone marrow mesenchymal stem cells for treatment of knee osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(5):736-741. doi: 10.3969/j.issn.2095-4344.2017.05.014
[7]	尹路, 蒋川锋, 陈俊杰, 等. 沙苑子苷A对关节软骨细胞凋亡的影响[J]. 中国组织工程研究, 2025, 29(8):1541-1547. doi: 10.12307/2025.339
	Yin L, Jiang CF, Chen JJ, et al. Effect of complanatoside A on the apoptosis of articular chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(8):1541-1547. doi: 10.12307/2025.339
[8]	Lai NS, Yu HC, Tung CH, et al. The role of aberrant expression of T cell mirnas affected by TNF-α in the immunopathogenesis of rheumatoid arthritis[J]. Arthritis Res Ther, 2017, 19(1):261. doi: 10.1186/s13075-017-1465-z
[9]	Stanczyk J, Pedrioli DM, Brentano F, et al. Altered expression of microRNA in synovial fibroblasts and synovial tissue in rheumatoid arthritis[J]. Arthritis Rheum, 2008, 58(4):1001-1009. doi: 10.1002/art.23386
[10]	Huang J, Zhao L, Fan Y, et al. The microRNAs miR-204 and miR-211 maintain joint homeostasis and protect against osteoarthritis progression[J]. Nat Commun, 2019, 10(1):2876. doi: 10.1038/s41467-019-10753-5
[11]	Wang QS, Fan KJ, Teng H, et al. Mir204 and Mir211 suppress synovial inflammation and proliferation in rheumatoid arthritis by targeting Ssrp1[J]. Elife, 2022, 11: e78085. doi: 10.7554/eLife.78085
[12]	Mei B, Chen J, Yang N, et al. The regulatory mechanism and biological significance of the snail-miR590-VEGFR-NRP1 axis in the angiogenesis, growth and metastasis of gastric cancer[J]. Cell Death Dis, 2020, 11(4):241. doi: 10.1038/s41419-020-2428-x
[13]	Wang L, Zhang Z, Yu X, et al. SOX9/miR-203a axis drives PI3K/AKT signaling to promote esophageal cancer progression[J]. Cancer Lett, 2020, 468: 14-26. doi: 10.1016/j.canlet.2019.10.004
[14]	Hardy RS, Hülso C, Liu Y, et al. Characterisation of fibroblast-like synoviocytes from a murine model of joint inflammation[J]. Arthritis Res Ther, 2013, 15(1):R24. doi: 10.1186/ar4158
[15]	Sendo S, Machado CRL, Boyle DL, et al. Dysregulated NUB1 and neddylation enhances rheumatoid arthritis fibroblast-like synoviocyte inflammatory responses[J]. Arthritis Rheumatol, 2024, 76(8):1252-1262. doi: 10.1002/art.42856
[16]	Wu J, Zhao FT, Fan KJ, et al. Dihydromyricetin inhibits inflammation of fibroblast-like synoviocytes through regulation of nuclear factor- κB signaling in rats with collagen-induced arthritis[J]. J Pharmacol Exp Ther, 2019, 368(2):218-228. doi: 10.1124/jpet.118.253369
[17]	王琪珊, 范凯健, 许冰馨, 等. DLTH通过调节免疫功能和软骨降解从而抑制RA的作用[J]. 中国药理学通报, 2021, 37(5):624-630. doi: 10.3969/j.issn.1001-1978.2021.05.007
	Wang QS, Fan KJ, Xu BX, et al. Study of DLTH inhibiting RA by regulating immune function and cartilage degradation[J]. Chinese Pharmacological Bulletin, 2021, 37(5):624-630. doi: 10.3969/j.issn.1001-1978.2021.05.007
[18]	伍沙沙, 王延, 徐婷, 等. 成纤维样滑膜细胞在类风湿关节炎发病机制中的作用[J]. 风湿病与关节炎, 2022, 11(2):43-47. doi: 10.7619/jcmp.20221368
	Wu SS, Wang Y, Xu T, et al. Role of fibroblast-like synoviocytes in the pathogenesis of rheumatoid arthritis[J]. Rheumatism and Arthritis, 2022, 11(2):43-47. doi: 10.7619/jcmp.20221368
[19]	Zhao L, Wu Q, Long Y, et al. microRNAs: critical targets for treating rheumatoid arthritis angiogenesis[J]. J Drug Target, 2024, 32(1):1-20. doi: 10.1080/1061186X.2023.2284097
[20]	Kmiołek T, Paradowska-Gorycka A. miRNAs as biomarkers and possible therapeutic strategies in rheumatoid arthritis[J]. Cells, 2022, 11(3):452. doi: 10.3390/cells11030452
[21]	Ouyang Y, Wang W, Tu B, et al. Overexpression of SOX9 alleviates the progression of human osteoarthritis in vitro and in vivo[J]. Drug Des Devel Ther, 2019, 13: 2833-2842. doi: 10.2147/DDDT.S203974
[22]	Jouan Y, Bouchemla Z, Bardèche-Trystram B, et al. Lin28a induces SOX9 and chondrocyte reprogramming via HMGA2 and blunts cartilage loss in mice[J]. Sci Adv, 2022, 8(34):eabn3106. doi: 10.1126/sciadv.abn3106
[23]	Shi C, Zheng W, Wang J. lncRNA-CRNDE regulates BMSC chondrogenic differentiation and promotes cartilage repair in osteoarthritis through SIRT1/SOX9[J]. Mol Cell Biochem, 2021, 476(4):1881-1890. doi: 10.1007/s11010-020-04047-4
[24]	Feng C, Ma F, Hu C, et al. SOX9/miR-130a/CTR1 axis modulates DDP-resistance of cervical cancer cell[J]. Cell Cycle, 2018, 17(4):448-458. doi: 10.1080/15384101.2017.1395533
[25]	Shi S, Wang C, Acton AJ, et al. Role of sox9 in growth factor regulation of articular chondrocytes[J]. J Cell Biochem, 2015, 116(7):1391-1400. doi: 10.1002/jcb.25099

组别	Sox9	P
siNC	1.00 ± 0.10
siSox9-1	0.70 ± 0.11	< 0.01
siSox9-2	0.66 ± 0.08	< 0.001
siSox9-3	0.50 ± 0.10	< 0.0001

组别	P16	P21	P53
siNC	1.00 ± 0.12	1.00 ± 0.21	1.00 ± 0.22
siSox9	1.95 ± 0.34	1.64 ± 0.19	1.98 ± 0.51