Upregulating miR‑27a‑3p inhibits cell proliferation and inflammation of rheumatoid arthritis synovial fibroblasts through targeting toll‑like receptor 5

Chen,Lifeng; Lu,Qiping; Chen,Jianhua; Feng,Ruibing; Yang,Chenxi

doi:10.3892/etm.2021.10661

Upregulating miR‑27a‑3p inhibits cell proliferation and inflammation of rheumatoid arthritis synovial fibroblasts through targeting toll‑like receptor 5

Authors:
- Lifeng Chen
- Qiping Lu
- Jianhua Chen
- Ruibing Feng
- Chenxi Yang
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Affiliations: Department of Rheumatology and Immunology, General Hospital of Central Theater Command, Wuhan, Hubei 430070, P.R. China, Department of General Surgery, General Hospital of Central Theater Command, Wuhan, Hubei 430070, P.R. China, Department of Orthopedics, Central People's Liberation Army Central Theater, Wuhan, Hubei 430070, P.R. China, Department of Orthopedics, Graduate School of Hubei University of Traditional Chinese Medicine, Wuhan, Hubei 430061, P.R. China
Published online on: August 27, 2021 https://doi.org/10.3892/etm.2021.10661
Article Number: 1227
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Rheumatoid arthritis (RA) is a serious chronic inflammatory disease and synovial fibroblasts (SFs) serve a vital role in the pathogenesis and progression of RA. Current studies have demonstrated that dysregulation of microRNAs is involved in RA etiopathogenesis. The present study aimed to investigate the role of microRNA (miR)‑27a‑3p in RASFs, as well as its molecular mechanism. RASFs were isolated from synovial tissues from patients with RA. Expression of miR‑27a‑3p and toll‑like receptor 5 (TLR5) was detected using reverse transcription‑quantitative polymerase chain reaction and western blotting. Cell proliferation, apoptosis and inflammatory response were measured with MTT assay, flow cytometry and ELISA kits, respectively. The target binding between miR‑27a‑3p and TLR5 was predicted on DIANA TOOLS software, and confirmed by dual‑luciferase reporter assay and Biotin‑coupled miRNA pull‑down assay. Expression of miR‑27a‑3p was downregulated and TLR5 was upregulated in synovial tissues and RASFs isolated from patients with RA. Functionally, upregulating miR‑27a‑3p may promote the apoptosis rate of RASFs and suppress cell proliferation and secretions of interleukin (IL)‑1β, IL‑6 and tumor necrosis factor‑α. TLR5 was validated as a downstream target for miR‑27a‑3p in RASFs, and its expression was negatively regulated by miR‑27a‑3p. Silencing TLR5 in RASFs may exert similar effects to miR‑27a‑3p‑overexpression; whereas, restoring TLR5 counteracted the suppression of miR‑27a‑3p‑overexpression on RASF proliferation and inflammation, as well as the promotion on apoptosis. miR‑27a‑3p upregulation may suppress RA progression by inhibiting RASFs proliferation and inflammation through targeting TLR5.

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1	Coras R, Narasimhan R and Guma M: Liquid biopsies to guide therapeutic decisions in rheumatoid arthritis. Transl Res. 201:1–12. 2018.PubMed/NCBI View Article : Google Scholar
2	Araki Y and Mimura T: The mechanisms underlying chronic inflammation in rheumatoid arthritis from the perspective of the epigenetic landscape. J Immunol Res. 2016(6290682)2016.PubMed/NCBI View Article : Google Scholar
3	Croia C, Bursi R, Sutera D, Petrelli F, Alunno A and Puxeddu I: One year in review 2019: Pathogenesis of rheumatoid arthritis. Clin Exp Rheumatol. 37:347–357. 2019.PubMed/NCBI
4	Fang Q, Zhou C and Nandakumar KS: Molecular and cellular pathways contributing to joint damage in rheumatoid arthritis. Mediators Inflamm. 2020(3830212)2020.PubMed/NCBI View Article : Google Scholar
5	Alunno A, Carubbi F, Giacomelli R and Gerli R: Cytokines in the pathogenesis of rheumatoid arthritis: New players and therapeutic targets. BMC Rheumatol. 1(3)2017.PubMed/NCBI View Article : Google Scholar
6	Falconer J, Murphy AN, Young SP, Clark AR, Tiziani S, Guma M and Buckley CD: Review: Synovial cell metabolism and chronic inflammation in rheumatoid arthritis. Arthritis Rheumatol. 70:984–999. 2018.PubMed/NCBI View Article : Google Scholar
7	Shi J, Ermann J, Weissman BN, Smith SE and Mandell JC: Thinking beyond pannus: A review of retro-odontoid pseudotumor due to rheumatoid and non-rheumatoid etiologies. Skeletal Radiol. 48:1511–1523. 2019.PubMed/NCBI View Article : Google Scholar
8	Laev SS and Salakhutdinov NF: Anti-arthritic agents: Progress and potential. Bioorg Med Chem. 23:3059–3080. 2015.PubMed/NCBI View Article : Google Scholar
9	Yuasa S, Yamaguchi H, Nakanishi Y, Kawaminami S, Tabata R, Shimizu N, Kohno M, Shimizu T, Miyata J, Nakayama M, et al: Treatment responses and their predictors in patients with rheumatoid arthritis treated with biological agents. J Med Invest. 60:77–90. 2013.PubMed/NCBI View Article : Google Scholar
10	Zamanpoor M: The genetic pathogenesis, diagnosis and therapeutic insight of rheumatoid arthritis. Clin Genet. 95:547–557. 2019.PubMed/NCBI View Article : Google Scholar
11	Moran-Moguel MC, Petarra-Del Rio S, Mayorquin-Galvan EE and Zavala-Cerna MG: Rheumatoid arthritis and mirnas: A critical review through a functional view. J Immunol Res. 2018(2474529)2018.PubMed/NCBI View Article : Google Scholar
12	Dong H, Lei J, Ding L, Wen Y, Ju H and Zhang X: MicroRNA: Function, detection, and bioanalysis. Chem Rev. 113:6207–6233. 2013.PubMed/NCBI View Article : Google Scholar
13	Dudics S, Venkatesha SH and Moudgil KD: The micro-RNA expression profiles of autoimmune arthritis reveal novel biomarkers of the disease and therapeutic response. Int J Mol Sci. 19(19)2018.PubMed/NCBI View Article : Google Scholar
14	Zhang H, Huang X, Ye L, Guo G, Li X, Chen C, Sun L, Li B, Chen N and Xue X: B cell-related circulating MicroRNAs with the potential value of biomarkers in the differential diagnosis, and distinguishment between the disease activity and lupus nephritis for systemic lupus erythematosus. Front Immunol. 9(1473)2018.PubMed/NCBI View Article : Google Scholar
15	Regev K, Paul A, Healy B, von Glenn F, Diaz-Cruz C, Gholipour T, Mazzola MA, Raheja R, Nejad P, Glanz BI, et al: Comprehensive evaluation of serum microRNAs as biomarkers in multiple sclerosis. Neurol Neuroimmunol Neuroinflamm. 3(e267)2016.PubMed/NCBI View Article : Google Scholar
16	Evangelatos G, Fragoulis GE, Koulouri V and Lambrou GI: MicroRNAs in rheumatoid arthritis: From pathogenesis to clinical impact. Autoimmun Rev. 18(102391)2019.PubMed/NCBI View Article : Google Scholar
17	Sharma AR, Sharma G, Lee SS and Chakraborty C: miRNA-regulated key components of cytokine signaling pathways and inflammation in rheumatoid arthritis. Med Res Rev. 36:425–439. 2016.PubMed/NCBI View Article : Google Scholar
18	Sode J, Krintel SB, Carlsen AL, Hetland ML, Johansen JS, Hørslev-Petersen K, Stengaard-Pedersen K, Ellingsen T, Burton M, Junker P, et al: Plasma MicroRNA profiles in patients with early rheumatoid arthritis responding to adalimumab plus methotrexate vs, methotrexate alone: A Placebo-controlled Clinical Trial. J Rheumatol. 45:53–61. 2018.PubMed/NCBI View Article : Google Scholar
19	Takamura Y, Aoki W, Satomura A, Shibasaki S and Ueda M: Small RNAs detected in exosomes derived from the MH7A synovial fibroblast cell line with TNF-α stimulation. PLoS One. 13(e0201851)2018.PubMed/NCBI View Article : Google Scholar
20	Maeda Y, Farina NH, Matzelle MM, Fanning PJ, Lian JB and Gravallese EM: Synovium-derived MicroRNAs regulate bone pathways in rheumatoid arthritis. J Bone Miner Res. 32:461–472. 2017.PubMed/NCBI View Article : Google Scholar
21	Shi DL, Shi GR, Xie J, Du XZ and Yang H: MicroRNA-27a inhibits cell migration and invasion of fibroblast-like synoviocytes by targeting follistatin-like protein 1 in rheumatoid arthritis. Mol Cells. 39:611–618. 2016.PubMed/NCBI View Article : Google Scholar
22	Aletaha D, Neogi T, Silman AJ, Funovits J, Felson DT, Bingham CO III, Birnbaum NS, Burmester GR, Bykerk VP, Cohen MD, et al: 2010 rheumatoid arthritis classification criteria: An American College of Rheumatology/European League Against Rheumatism collaborative initiative. Ann Rheum Dis. 69:1580–1588. 2010.PubMed/NCBI View Article : Google Scholar
23	Li XJ, Xu M, Zhao XQ, Zhao JN, Chen FF, Yu W, Gao DY and Luo B: Proteomic analysis of synovial fibroblast-like synoviocytes from rheumatoid arthritis. Clin Exp Rheumatol. 31:552–558. 2013.PubMed/NCBI
24	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
25	Zou Y, Xu S, Xiao Y, Qiu Q, Shi M, Wang J, Liang L, Zhan Z, Yang X, Olsen N, et al: Long noncoding RNA LERFS negatively regulates rheumatoid synovial aggression and proliferation. J Clin Invest. 128:4510–4524. 2018.PubMed/NCBI View Article : Google Scholar
26	Alamanos Y and Drosos AA: Epidemiology of adult rheumatoid arthritis. Autoimmun Rev. 4:130–136. 2005.PubMed/NCBI View Article : Google Scholar
27	Liu Y, Pan YF, Xue YQ, Fang LK, Guo XH, Guo X, Liu M, Mo BY, Yang MR, Liu F, et al: uPAR promotes tumor-like biologic behaviors of fibroblast-like synoviocytes through PI3K/Akt signaling pathway in patients with rheumatoid arthritis. Cell Mol Immunol. 15:171–181. 2018.PubMed/NCBI View Article : Google Scholar
28	Cai P, Jiang T, Li B, Qin X, Lu Z, Le Y, Shen C, Yang Y, Zheng L and Zhao J: Comparison of rheumatoid arthritis (RA) and osteoarthritis (OA) based on microarray profiles of human joint fibroblast-like synoviocytes. Cell Biochem Funct. 37:31–41. 2019.PubMed/NCBI View Article : Google Scholar
29	Ye Y, Gao X and Yang N: LncRNA ZFAS1 promotes cell migration and invasion of fibroblast-like synoviocytes by suppression of miR-27a in rheumatoid arthritis. Hum Cell. 31:14–21. 2018.PubMed/NCBI View Article : Google Scholar
30	Yang S and Yang Y: Downregulation of microRNA 221 decreases migration and invasion in fibroblast like synoviocytes in rheumatoid arthritis. Mol Med Rep. 12:2395–2401. 2015.PubMed/NCBI View Article : Google Scholar
31	Neumann E, Lefèvre S, Zimmermann B, Gay S and Müller-Ladner U: Rheumatoid arthritis progression mediated by activated synovial fibroblasts. Trends Mol Med. 16:458–468. 2010.PubMed/NCBI View Article : Google Scholar
32	Karouzakis E, Gay RE, Gay S and Neidhart M: Epigenetic control in rheumatoid arthritis synovial fibroblasts. Nat Rev Rheumatol. 5:266–272. 2009.PubMed/NCBI View Article : Google Scholar
33	Li H, Guan SB, Lu Y and Wang F: MiR-140-5p inhibits synovial fibroblasts proliferation and inflammatory cytokines secretion through targeting TLR4. Biomed Pharmacother. 96:208–214. 2017.PubMed/NCBI View Article : Google Scholar
34	Fu D, Xiao C, Xie Y, Gao J and Ye S: MiR-3926 inhibits synovial fibroblasts proliferation and inflammatory cytokines secretion through targeting toll like receptor 5. Gene. 687:200–206. 2019.PubMed/NCBI View Article : Google Scholar
35	Thunyakitpisal P, Ruangpornvisuti V, Kengkwasing P, Chokboribal J and Sangvanich P: Acemannan increases NF-κB/DNA binding and IL-6/-8 expression by selectively binding Toll-like receptor-5 in human gingival fibroblasts. Carbohydr Polym. 161:149–157. 2017.PubMed/NCBI View Article : Google Scholar
36	Sabroe I, Read RC, Whyte MK, Dockrell DH, Vogel SN and Dower SK: Toll-like receptors in health and disease: Complex questions remain. J Immunol. 171:1630–1635. 2003.PubMed/NCBI View Article : Google Scholar
37	Elshabrawy HA, Essani AE, Szekanecz Z, Fox DA and Shahrara S: TLRs, future potential therapeutic targets for RA. Autoimmun Rev. 16:103–113. 2017.PubMed/NCBI View Article : Google Scholar
38	Thwaites RS, Unterberger S, Chamberlain G, Walker-Bone K, Davies KA and Sacre S: TLR1/2 and 5 induce elevated cytokine levels from rheumatoid arthritis monocytes independent of ACPA or RF autoantibody status. Rheumatology (Oxford). 59:3533–3539. 2020.PubMed/NCBI View Article : Google Scholar
39	Smith KD, Andersen-Nissen E, Hayashi F, Strobe K, Bergman MA, Barrett SL, Cookson BT and Aderem A: Toll-like receptor 5 recognizes a conserved site on flagellin required for protofilament formation and bacterial motility. Nat Immunol. 4:1247–1253. 2003.PubMed/NCBI View Article : Google Scholar
40	Wu YW, Tang W and Zuo JP: Toll-like receptors: Potential targets for lupus treatment. Acta Pharmacol Sin. 36:1395–1407. 2015.PubMed/NCBI View Article : Google Scholar
41	Chamberlain ND, Vila OM, Volin MV, Volin MV, Volkov S, Pope RM, Swedler W, Mandelin AM 2nd, Shahrara S, et al: TLR5, a novel and unidentified inflammatory mediator in rheumatoid arthritis that correlates with disease activity score and joint TNF-α levels. J Immunol. 189:475–483. 2012.PubMed/NCBI View Article : Google Scholar
42	Kim SJ, Chen Z, Chamberlain ND, Essani AB, Volin MV, Amin MA, Volkov S, Gravallese EM, Arami S, Swedler W, et al: Ligation of TLR5 promotes myeloid cell infiltration and differentiation into mature osteoclasts in rheumatoid arthritis and experimental arthritis. J Immunol. 193:3902–3913. 2014.PubMed/NCBI View Article : Google Scholar