Transforming growth factor β1 promotes fibroblast-like synoviocytes migration and invasion via TGF-β1/Smad signaling in rheumatoid arthritis
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Migration and invasion are important characteristics of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs), which are involved in joint damage and contribute to rheumatoid arthritis (RA) pathology. However, the underlying mechanisms remain unclear. Because epithelial–mesenchymal transition (EMT) is a key mechanism related to migration and invasion in cancer cells, we investigated the relationship between EMT and RA-FLSs and explored whether the transforming growth factor β1 (TGF-β1)/Smad signaling pathway is involved. In vivo, fibroblast-like synoviocytes (FLSs) were isolated from the synovium of RA or osteoarthritis (OA) patients and cultured for 4–8 passages. EMT markers were detected by immunofluorescence and Western blotting. RA-FLSs were treated with TGF-β1 or Smad2/3 small interfering RNA (siRNA), EMT markers were detected, and migration and invasion were assessed by Transwell assays. EMT markers could be detected in FLSs; when compared with osteoarthritis fibroblast-like synoviocytes (OA-FLSs), E-cadherin and vimentin decreased, while N-cadherin and α-smooth muscle actin (α-SMA) increased in RA-FLSs. Furthermore, TGF-β1 enhanced migration and invasion by inducing EMT via activating Smad2/3 in RA-FLSs. Phosphorylation of Smad2/3 was accompanied by degradation of Smad3. Silencing Smad2/3 blocked EMT and inhibited the migration and invasion induced by TGF-β1. Matrix metalloproteinase 9 (MMP9) and vimentin were not affected when cells were treated with TGF-β1 or Smad2/3 siRNA. The TGF-β1/Smad signaling pathway is involved in EMT and contributes to migration and invasion in RA-FLSs. Interestingly, vimentin decreased in RA-FLSs, but there is no correlation between vimentin and TGF-β1/Smad signaling pathway. Thus, further research on vimentin should be conducted.
KeywordsRheumatoid arthritis Epithelial–mesenchymal transition Transforming growth factor β1 Migration Invasion
The present study was supported by National Natural Science Foundation of China (Grant Nos. 81771747, 81801624) and Natural Science Foundation of Guangdong Province (Grant No. 2017A030313475).
Compliance with ethical standards
Conflict of interest
The authors have declared that no conflicts of interest exist.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethics Committee of Southern Medical University and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Additional informed consent was obtained from all individual participants for whom identifying information is included in this article.
- 7.Nakamura H, Shimamura S, Yasuda S, Kono M, Kono M, Fujieda Y, Kato M, Oku K, Bohgaki T, Shimizu T, Iwasaki N, Atsumi T (2018) Ectopic RASGRP2 (CalDAG-GEFI) expression in rheumatoid synovium contributes to the development of destructive arthritis. Ann Rheum Dis 77(12):1765–1772. https://doi.org/10.1136/annrheumdis-2018-213588 CrossRefGoogle Scholar
- 9.Lefèvre S, Knedla A, Tennie C, Kampmann A, Wunrau C, Dinser R, Korb A, Schnäker E, Tarner IH, Robbins PD, Evans CH, Stürz H, Steinmeyer J, Gay S, Schölmerich J, Pap T, Müller-Ladner U, Neumann E (2009) Synovial fibroblasts spread rheumatoid arthritis to unaffected joints. Nat Med 15(12):1414–1420. https://doi.org/10.1038/nm.2050 CrossRefGoogle Scholar
- 14.Tian X, Guan W, Zhang L, Sun W, Zhou D, Lin Q, Ren W, Nadeem L, Xu G (2018) Physical interaction of STAT1 isoforms with TGF-β receptors leads to functional crosstalk between two signaling pathways in epithelial ovarian cancer. J Exp Clin Cancer Res 37(1):103. https://doi.org/10.1186/s13046-018-0773-8 CrossRefGoogle Scholar
- 16.Song HY, Kim MY, Kim KH, Lee IH, Shin SH, Lee JS, Kim JH (2010) Synovial fluid of patients with rheumatoid arthritis induces α-smooth muscle actin in human adipose tissue-derived mesenchymal stem cells through a TGF-β1-dependent mechanism. Exp Mol Med 42(8):565. https://doi.org/10.3858/emm.2010.42.8.057 CrossRefGoogle Scholar
- 18.Li G, Zhang Y, Liu D, Qian Y, Zhang H, Guo S, Sunagawa M, Hisamitsu T, Liu Y (2013) PI3 kinase/Akt/HIF-1α pathway is associated with hypoxia-induced epithelial–mesenchymal transition in fibroblast-like synoviocytes of rheumatoid arthritis. Mol Cell Biochem 372(1–2):221–231. https://doi.org/10.1007/s11010-012-1463-z CrossRefGoogle Scholar
- 20.Steenvoorden MM, Tolboom TC, van der Pluijm G, Löwik C, Visser CP, DeGroot J, Gittenberger-DeGroot AC, DeRuiter MC, Wisse BJ, Huizinga TW, Toes RE (2006) Transition of healthy to diseased synovial tissue in rheumatoid arthritis is associated with gain of mesenchymal/fibrotic characteristics. Arthritis Res Ther 8(6):R165. https://doi.org/10.1186/ar2073 CrossRefGoogle Scholar
- 21.Dhawan U, Sue M, Lan K, Buddhakosai W, Huang PH, Chen YC, Chen P, Chen WL (2018) Nanochip-induced epithelial-to-mesenchymal transition: impact of physical microenvironment on cancer metastasis. ACS Appl Mater Interfaces 10(14):11474–11485. https://doi.org/10.1021/acsami.7b19467 CrossRefGoogle Scholar
- 22.Wang J, Guan X, Zhang Y, Ge S, Zhang L, Li H, Wang X, Liu R, Ning T, Deng T, Zhang H, Jiang X, Ba Y, Huang D (2018) Exosomal miR-27a derived from gastric cancer cells regulates the transformation of fibroblasts into cancer-associated fibroblasts. Cell Physiol Biochem 49(3):869–883. https://doi.org/10.1159/000493218 CrossRefGoogle Scholar
- 25.Filer A, Ward LSC, Kemble S, Davies CS, Munir H, Rogers R, Raza K, Buckley CD, Nash GB, McGettrick HM (2017) Identification of a transitional fibroblast function in very early rheumatoid arthritis. Ann Rheum Dis 76(12):2105–2112. https://doi.org/10.1136/annrheumdis-2017-211286 CrossRefGoogle Scholar
- 27.Gonzalo-Gil E, Criado G, Santiago B, Dotor J, Pablos JL, Galindo M (2013) Transforming growth factor (TGF)-beta signalling is increased in rheumatoid synovium but TGF-beta blockade does not modify experimental arthritis. Clin Exp Immunol 174(2):245–255. https://doi.org/10.1111/cei.12179 Google Scholar
- 29.Xiao L, Peng X, Liu F, Tang C, Hu C, Xu X, Wang M, Luo Y, Yang S, Song P, Xiao P, Kanwar YS, Sun L (2015) AKT regulation of mesothelial-to-mesenchymal transition in peritoneal dialysis is modulated by smurf2 and deubiquitinating enzyme USP4. BMC Cell Biol 16(1):7. https://doi.org/10.1186/s12860-015-0055-7 CrossRefGoogle Scholar
- 32.Kim ES, Kim MS, Moon A (2004) TGF-beta-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK, but not ERK signaling in MCF10A human breast epithelial cells. Int J Oncol 25(5):1375–1382Google Scholar
- 34.Okamoto T, Takahashi S, Nakamura E, Nagaya K, Hayashi T, Fujieda K (2009) Transforming growth factor-beta1 induces matrix metalloproteinase-9 expression in human meningeal cells via ERK and Smad pathways. Biochem Biophys Res Commun 383(4):475–479. https://doi.org/10.1016/j.bbrc.2009.04.038 CrossRefGoogle Scholar
- 35.Etienne-Manneville S (2018) Cytoplasmic intermediate filaments in cell biology. Annu Rev Cell Dev Biol 34(1):1–28. https://doi.org/10.1146/annurev-cellbio-100617-062534 CrossRefGoogle Scholar
- 37.Dmello C, Sawant S, Alam H, Gangadaran P, Tiwari R, Dongre H, Rana N, Barve S, Costea DE, Chaukar D, Kane S, Pant H, Vaidya M (2016) Vimentin-mediated regulation of cell motility through modulation of beta4 integrin protein levels in oral tumor derived cells. Int J Biochem Cell Biol 70:161–172. https://doi.org/10.1016/j.biocel.2015.11.015 CrossRefGoogle Scholar
- 39.Fan LY, He DY, Wang Q, Zong M, Zhang H, Yang L, Sun LS (2012) Citrullinated vimentin stimulates proliferation, pro-inflammatory cytokine secretion, and PADI4 and RANKL expression of fibroblast-like synoviocytes in rheumatoid arthritis. Scand J Rheumatol 41(5):354–358. https://doi.org/10.3109/03009742.2012.670263 CrossRefGoogle Scholar