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Transforming growth factor β1 promotes fibroblast-like synoviocytes migration and invasion via TGF-β1/Smad signaling in rheumatoid arthritis

  • DingJi Zhu
  • JinJun Zhao
  • AiJu Lou
  • Qin Huang
  • QingQing OuYang
  • JunQing Zhu
  • MeiDa Fan
  • YingQiong He
  • Hao RenEmail author
  • Min YangEmail author
Article
  • 40 Downloads

Abstract

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.

Keywords

Rheumatoid arthritis Epithelial–mesenchymal transition Transforming growth factor β1 Migration Invasion 

Notes

Acknowledgements

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.

Ethical approval

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.

References

  1. 1.
    Klein K, Gay S (2013) Epigenetic modifications in rheumatoid arthritis, a review. Curr Opin Pharmacol 13(3):420–425.  https://doi.org/10.1016/j.coph.2013.01.007 CrossRefGoogle Scholar
  2. 2.
    Scott DL, Wolfe F, Huizinga TW (2010) Rheumatoid arthritis. Lancet 376(9746):1094–1108.  https://doi.org/10.1016/S0140-6736(10)60826-4 CrossRefGoogle Scholar
  3. 3.
    Kourilovitch M, Galarza-Maldonado C, Ortiz-Prado E (2014) Diagnosis and classification of rheumatoid arthritis. J Autoimmun 48–49:26–30.  https://doi.org/10.1016/j.jaut.2014.01.027 CrossRefGoogle Scholar
  4. 4.
    Smolen JS, Aletaha D, McInnes IB (2016) Rheumatoid arthritis. Lancet 388(10055):1984CrossRefGoogle Scholar
  5. 5.
    Carmona L, Cross M, Williams B, Lassere M, March L (2010) Rheumatoid arthritis. Best Pract Res Clin Rheumatol 24(6):733–745.  https://doi.org/10.1016/j.berh.2010.10.001 CrossRefGoogle Scholar
  6. 6.
    Klareskog L, Catrina AI, Paget S (2009) Rheumatoid arthritis. Lancet 373(9664):659–672.  https://doi.org/10.1016/S0140-6736(09)60008-8 CrossRefGoogle Scholar
  7. 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
  8. 8.
    Neumann E, Lefèvre S, Zimmermann B, Gay S, Müller-Ladner U (2010) Rheumatoid arthritis progression mediated by activated synovial fibroblasts. Trends Mol Med 16(10):458–468.  https://doi.org/10.1016/j.molmed.2010.07.004 CrossRefGoogle Scholar
  9. 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
  10. 10.
    Thiery JP, Acloque H, Huang RYJ, Nieto MA (2009) Epithelial–mesenchymal transitions in development and disease. Cell 139(5):871–890.  https://doi.org/10.1016/j.cell.2009.11.007 CrossRefGoogle Scholar
  11. 11.
    Shi L, Dong N, Fang X, Wang X (2016) Regulatory mechanisms of TGF-β1-induced fibrogenesis of human alveolar epithelial cells. J Cell Mol Med 20(11):2183–2193.  https://doi.org/10.1111/jcmm.12918 CrossRefGoogle Scholar
  12. 12.
    Zhang C, Hao Y, Wang Y, Xu J, Teng Y, Yang X (2018) TGF-β/SMAD4-regulated LncRNA-LINP1 inhibits epithelial–mesenchymal transition in lung cancer. Int J Biol Sci 14(12):1715–1723.  https://doi.org/10.7150/ijbs.27197 CrossRefGoogle Scholar
  13. 13.
    Wu X, Zhao J, Ruan Y, Sun L, Xu C, Jiang H (2018) Sialyltransferase ST3GAL1 promotes cell migration, invasion, and TGF-β1-induced EMT and confers paclitaxel resistance in ovarian cancer. Cell Death Dis 9(11):1102.  https://doi.org/10.1038/s41419-018-1101-0 CrossRefGoogle Scholar
  14. 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
  15. 15.
    Pohlers D, Beyer A, Koczan D, Wilhelm T, Thiesen H, Kinne RW (2007) Constitutive upregulation of the transforming growth factor-β pathway in rheumatoid arthritis synovial fibroblasts. Arthritis Res Ther 9(3):R59.  https://doi.org/10.1186/ar2217 CrossRefGoogle Scholar
  16. 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
  17. 17.
    Eliçabe RJ, Silva JE, Dave MN, Lacoste MG, Tamashiro H, Blas R, Munarriz A, Rabinovich GA, Di Genaro MS (2017) Association between IL-17 and IgA in the joints of patients with inflammatory arthropathies. BMC Immunol 18(1):8.  https://doi.org/10.1186/s12865-017-0189-9 CrossRefGoogle Scholar
  18. 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
  19. 19.
    Zvaifler NJ (2006) Relevance of the stroma and epithelial–mesenchymal transition (EMT) for the rheumatic diseases. Arthritis Res Ther 8(3):210.  https://doi.org/10.1186/ar1963 CrossRefGoogle Scholar
  20. 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. 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. 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
  23. 23.
    Chen S, Shiau A, Li Y, Lin C, Jou I, Liu M, Wu C, Wang C (2015) Transcription factor snail regulates tumor necrosis factor α-mediated synovial fibroblast activation in the rheumatoid joint. Arthritis Rheumatol 67(1):39–50.  https://doi.org/10.1002/art.38899 CrossRefGoogle Scholar
  24. 24.
    Giese G, Kubbies M, Traub P (1992) Cell cycle-dependent vimentin expression in elutriator-synchronized, TPA-treated MPC-11 mouse plasmacytoma cells. Exp Cell Res 200(1):118CrossRefGoogle Scholar
  25. 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
  26. 26.
    Massagué J (2012) TGFβ signalling in context. Nat Rev Mol Cell Biol 13(10):616–630.  https://doi.org/10.1038/nrm3434 CrossRefGoogle Scholar
  27. 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
  28. 28.
    Xu Z, Greenblatt MB, Yan G, Feng H, Sun J, Lotinun S, Brady N, Baron R, Glimcher LH, Zou W (2017) SMURF2 regulates bone homeostasis by disrupting SMAD3 interaction with vitamin D receptor in osteoblasts. Nat Commun 8:14570.  https://doi.org/10.1038/ncomms14570 CrossRefGoogle Scholar
  29. 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
  30. 30.
    Zhang Z, Finnerty CC, He J, Herndon DN (2012) Smad ubiquitination regulatory factor 2 expression is enhanced in hypertrophic scar fibroblasts from burned children. Burns 38(2):236–246.  https://doi.org/10.1016/j.burns.2011.08.012 CrossRefGoogle Scholar
  31. 31.
    Wang Y, Wan D, Zhou R, Zhong W, Lu S, Chai Y (2017) Geraniin inhibits migration and invasion of human osteosarcoma cancer cells through regulation of PI3K/Akt and ERK1/2 signaling pathways. Anti-cancer Drug 28(9):959–966.  https://doi.org/10.1097/CAD.0000000000000535 CrossRefGoogle Scholar
  32. 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
  33. 33.
    Hsieh HL, Wang HH, Wu WB, Chu PJ, Yang CM (2010) Transforming growth factor-beta1 induces matrix metalloproteinase-9 and cell migration in astrocytes: roles of ROS-dependent ERK- and JNK-NF-kappaB pathways. J Neuroinflammation 7:88.  https://doi.org/10.1186/1742-2094-7-88 CrossRefGoogle Scholar
  34. 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. 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
  36. 36.
    Leube RE, Moch M, Windoffer R (2015) Intermediate filaments and the regulation of focal adhesion. Curr Opin Cell Biol 32:13–20.  https://doi.org/10.1016/j.ceb.2014.09.011 CrossRefGoogle Scholar
  37. 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
  38. 38.
    Kim CW, Cho EH, Lee YJ, Kim YH, Hah Y, Kim DR (2006) Disease-specific proteins from rheumatoid arthritis patients. J Korean Med Sci 21(3):478.  https://doi.org/10.3346/jkms.2006.21.3.478 CrossRefGoogle Scholar
  39. 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
  40. 40.
    Connor AM, Mahomed N, Gandhi R, Keystone EC, Berger SA (2012) TNFα modulates protein degradation pathways in rheumatoid arthritis synovial fibroblasts. Arthritis Res Ther 14(2):R62.  https://doi.org/10.1186/ar3778 CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Rheumatology and Immunology, Nanfang HospitalSouthern Medical UniversityGuangzhouPeople’s Republic of China
  2. 2.Department of Rheumatology and ImmunologyLiwan Hospital of The Third Affiliated Hospital, Guangzhou Medical UniversityGuangdongChina
  3. 3.Department of Ultrasound, The Fifth Affiliated HospitalSun Yat Sen UniversityGuangzhouChina

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