Inhibition of microRNA-21 decreases the invasiveness of fibroblast-like synoviocytes in rheumatoid arthritis via TGFβ/Smads signaling pathway

Document Type : Original Article

Authors

Department of Orthopedics, The First People’s Hospital of Hefei, Hefei, Anhui 230001, P.R. China.

Abstract

Objective(s): MicroRNA-21 (miR21) is aberrantly elevated in rheumatoid arthritis (RA) patients, the significance of this microRNA in RA pathogenesis and treatment, however, has not been investigated. In this study, by using RA-derived fibroblast-like synoviocyte (FLS) cells as a model, we investigated the effect and corresponding mechanism of miR21 inhibition on FLSs invasion.
Materials and Methods:miR21 expression in synovial tissue and FLSs in RA patients and non-RA controls were determined by stem-loop RT-PCR. The effect of miR21 on FLSs viability and invasiveness were evaluated using miR21 inhibition. Cell viability was evaluated by MTT assay and the expression of genes at mRNA and protein levels was determined by RT-PCR and Western blot, respectively.
Results: Our results showed that miR21 expression was highly increased in synovial tissue and FLSs in RA patients. Also, we reported that miR21 inhibitor treatment could significantly suppress the invasiveness of FLSs without affecting cell viability. The decreased FLSs invasion by miR21 inhibition was associated with down-regulated expression of matrix metalloproteinase (MMP)-1, MMP3, and MMP13. Further analysis revealed that miR21 inhibition could suppress the expression of TGFβ1 and Smad4, but promote that of Smad7. Moreover, suppression of FLS invasion and MMPs expression by miR21 treatment could be counteracted by additional TGFβ1 treatment.
Conclusion:Our results indicated that miR21 inhibition can down-regulate the expression of MMP1, MMP3, and MMP13 and consequently suppress the invasiveness of FLS, which is achieved through TGFβ1/Smad4/7 signaling pathway. The findings of this study could offer a novel approach for RA treatment.

Keywords

References

1. Klareskog L, Catrina AI, Paget S. Rheumatoid arthritis. Lancet 2009; 373:659-672.
2. Firestein GS. Evolving concepts of rheumatoid arthritis. Nature 2003; 423:356-361.
3. Matcham F, Ali S, Hotopf M, Chalder T. Psychological correlates of fatigue in rheumatoid arthritis: a systematic review. Clin Psychol Rev 2015; 39:16-29.
4. Mirfeizi Z, Noubakht Z, Rezaie AE, Jokar MH, Sarabi ZS. Plasma levels of leptin and visfatin in rheumatoid arthritis patients; is there any relationship with joint damage? Iran J Basic Med Sci 2014; 17:662-666.
5. Tolboom TC, van der Helm-Van Mil AH, Nelissen RG, Breedveld FC, Toes RE, Huizinga TW. Invasiveness of fibroblast-like synoviocytes is an individual patient characteristic associated with the rate of joint destruction in patients with rheumatoid arthritis. Arthritis Rheum 2005; 52:1999-2002.
6. Burrage PS, Mix KS, Brinckerhoff CE. Matrix metalloproteinases: role in arthritis. Front Biosci 2006; 11:529-543.
7. Tolboom TC, Pieterman E, van der Laan WH, Toes RE, Huidekoper AL, Nelissen RG, et al. Invasive properties of fibroblast-like synoviocytes: correlation with growth characteristics and expression of MMP-1, MMP-3, and MMP-10. Ann Rheum Dis 2002; 61:975-980.
8. Chen WX, Ren LH, Shi RH. Implication of miRNAs for inflammatory bowel disease treatment: Systematic review. World J Gastrointest Pathophysiol 2014; 5:63-70.
9.  Chatzikyriakidou A, Voulgari PV, Georgiou I, Drosos AA. miRNAs and related polymorphisms in rheumatoid arthritis susceptibility. Autoimmun Rev 2012; 11:636-641.
10. Latruffe N, Lancon A, Frazzi R, Aires V, Delmas D, Michaille JJ, et al. Exploring new ways of regulation by resveratrol involving miRNAs, with emphasis on inflammation. Ann N Y Acad Sci 2015; 1348:97-106.
11. Zhang Y, Pan T, Zhong X, Cheng C. Nicotine upregulates microRNA-21 and promotes TGF-beta-dependent epithelial-mesenchymal transition of esophageal cancer cells. Tumour Biol 2014; 35:7063-7072.
12. Cottonham CL, Kaneko S, Xu L. miR-21 and miR-31 converge on TIAM1 to regulate migration and invasion of colon carcinoma cells. J Biol Chem 2010; 285:35293-35302.
13. Alves C, Luime JJ, van Zeben D, Huisman A-M, Weel AEAM, Barendregt PJ, et al. Diagnostic performance of the ACR/EULAR 2010 criteria for rheumatoid arthritis and two diagnostic algorithms in an early arthritis clinic (REACH). Ann Rheum Dis 2011; 70:1645-1647.
14. Association WM. Declaration of Helsinki. Ethical principles for medical research involving human subjects. 2009.
15. Harada S, Yamamura M, Okamoto H, Morita Y, Kawashima M, Aita T, et al. Production of interleukin-7 and interleukin-15 by fibroblast-like synoviocytes from patients with rheumatoid arthritis. Arthritis Rheum 1999; 42:1508-1516.
16. Yan L-X, Huang X-F, Shao Q, Huang MAY, Deng L, Wu Q-L, et al. MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. RNA 2008; 14:2348-2360.
17. Stockert JC, Blázquez-Castro A, Cañete M, Horobin RW, Villanueva Á. MTT assay for cell viability: Intracellular localization of the formazan product is in lipid droplets. Acta Histochem 2012; 114:785-796.
18. Shridhar R, Zhang J, Song J, Booth BA, Kevil CG, Sotiropoulou G, et al. Cystatin M suppresses the malignant phenotype of human MDA-MB-435S cells. Oncogene 2004; 23:2206-2215.
19. Shi Y, Hata A, Lo RS, Massague J, Pavletich NP. A structural basis for mutational inactivation of the tumour suppressor Smad4. Nature 1997; 388:87-93.
20. Itoh F, Asao H, Sugamura K, Heldin CH, ten Dijke P, Itoh S. Promoting bone morphogenetic protein signaling through negative regulation of inhibitory Smads. EMBO J 2001; 20:4132-4142.
21. Saag KG, Teng GG, Patkar NM, Anuntiyo J, Finney C, Curtis JR, et al. American College of Rheumatology
2008 recommendations for the use of nonbiologic and biologic  disease-modifying antirheumatic drugs in rheumatoid arthritis. Arthritis Rheum 2008; 59:762-784.
22. Broderick JA, Zamore PD. MicroRNA therapeutics. Gene Ther 2011; 18:1104-1110.
23. Dangwal S, Thum T. microRNA therapeutics in cardiovascular disease models. Annu Rev Pharm Toxicol 2014; 54:185-203.
24. van Rooij E, Kauppinen S. Development of microRNA therapeutics is coming of age. EMBO Mol Med 2014; 6:851-864.
25. Thum T. MicroRNA therapeutics in cardiovascular medicine. EMBO Mol Med 2012; 4:3-14.
26. Wang H, Peng W, Ouyang X, Li W, Dai Y. Circulating microRNAs as candidate biomarkers in patients with systemic lupus erythematosus. Transl Res 2012; 160:198-206.
Iranian Journal of Basic Medical Sciences
Volume 19, Issue 7
July 2016
Pages 787-793

Files

Share

How to cite

Statistics