The Role of Focal Adhesion Kinase in the TGF-β-Induced Myofibroblast Transdifferentiation of Human Tenon’s Fibroblasts
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Purpose: To investigate the role of focal adhesion kinase (FAK) in transforming growth factor (TGF)-β-induced myofibroblast transdifferentiation of human Tenon’s fibroblasts. Methods: Primary cultured human Tenon’s fibroblasts were exposed to TGF-β1 for up to 48 hours. The mRNA levels of FAK, α smooth muscle actin (αSMA), and β-actin were determined by quantitative real time reverse transcription polymerase chain reaction. The protein levels of collagen type I, FAK, phospho-FAK, αSMA, and β-actin were determined by Western immunoblots. After the small interfering RNA targeting FAK (siRNAFAK) molecules were delivered into the cells, the expressions of αSMA proteins were determined by Western immunoblots. Results: In human Tenon’s fibroblasts, TGF-β1 significantly increased the mRNA and protein expressions of αSMA. However, when the action of FAK was inhibited using siRNAFAK, the TGF-β1-induced expression of αSMA was attenuated. Conclusions: Our data suggest that FAK may be associated with the TGF-β1-induced transdifferentiation of human Tenon’s fibroblasts to myofibroblasts, which is the essential step of subconjunctival fibrosis.
Abstract
Purpose: To investigate the role of focal adhesion kinase (FAK) in transforming growth factor (TGF)-β-induced myofibroblast transdifferentiation of human Tenon’s fibroblasts. Methods: Primary cultured human Tenon’s fibroblasts were exposed to TGF-β1 for up to 48 hours. The mRNA levels of FAK, α smooth muscle actin (αSMA), and β-actin were determined by quantitative real time reverse transcription polymerase chain reaction. The protein levels of collagen type I, FAK, phospho-FAK, αSMA, and β-actin were determined by Western immunoblots. After the small interfering RNA targeting FAK (siRNAFAK) molecules were delivered into the cells, the expressions of αSMA proteins were determined by Western immunoblots. Results: In human Tenon’s fibroblasts, TGF-β1 significantly increased the mRNA and protein expressions of αSMA. However, when the action of FAK was inhibited using siRNAFAK, the TGF-β1-induced expression of αSMA was attenuated. Conclusions: Our data suggest that FAK may be associated with the TGF-β1-induced transdifferentiation of human Tenon’s fibroblasts to myofibroblasts, which is the essential step of subconjunctival fibrosis.
참고문헌 (25)
- Eyden B, "The myofibroblast: an assessment of controversial issues and a definition useful in diagnosis and research", Ultrastruct Pathol, 25, pp.39~50 (2001)
- Eyden B, "Electron microscopy in the study of myofibroblastic lesions", Semin Diagn Pathol, 20, pp.13~24 (2003)
- Desmouliere A, "The stroma reaction myofibroblast: a key player in the control of tumor cell behavior", Int J Dev Biol, 48, pp.509~517 (2004)
- Meyer-Ter-Vehn T, "Localization of TGF-beta type II receptor and ED-A fibronectin in normal conjunctiva and failed filtering blebs", Mol Vis, 14, pp.136~141 (2008)
- Saika S, "TGF beta in fibroproliferative diseases in the eye", Front Biosci (Schol Ed), 1, pp.376~390 (2009)
- Seong GJ, "TGF-beta-induced interleukin-6 participates in transdifferentiation of human Tenon’s fibroblasts to myofibroblasts", Mol Vis, 15, pp.2123~2128 (2009)
- Nakamura H, "RNA interference targeting transforming growth factor-beta type II receptor suppresses ocular inflammation and fibrosis", Mol Vis, 10, pp.703~711 (2004)
- Meyer-Ter-Vehn T, "p38 inhibitors prevent TGF-beta-induced myofibroblast transdifferentiation in human tenon fibroblasts", Invest Ophthalmol Vis Sci, 47, pp.1500~1509 (2006)
- Yamanaka O, "Interleukin-7 modulates extracellular matrix production and TGF-beta signaling in cultured human subconjunctival fibroblasts", Curr Eye Res, 31, pp.491~499 (2006)
- Meyer-Ter-Vehn T, "Lovastatin inhibits TGF-beta-induced myofibroblast transdifferentiation in human tenon fibroblasts", Invest Ophthalmol Vis Sci, 49, pp.3955~3960 (2008)
- Chatzizacharias NA, "The role of focal adhesion kinase in early development", Histol Histopathol, 25, pp.1039~1055 (2010)
- Schwock J, "Targeting focal adhesion kinase signaling in tumor growth and metastasis", Expert Opin Ther Targets, 14, pp.77~94 (2010)
- Thannickal VJ, "Myofibroblast differentiation by transforming growth factor-beta1 is dependent on cell adhesion and integrin signaling via focal adhesion kinase", J Biol Chem, 278, pp.12384~12389 (2003)
- Mimura Y, "Constitutive phosphorylation of focal adhesion kinase is involved in the myofibroblast differentiation of scleroderma fibroblasts", J Invest Dermatol, 124, pp.886~892 (2005)
- Brenmoehl J, "Transforming growth factor-beta 1 induces intestinal myofibroblast differentiation and modulates their migration", World J Gastroenterol, 15, pp.1431~1442 (2009)
- McLean GW, "The role of focal-adhesion kinase in cancer: a new therapeutic opportunity", Nat Rev Cancer, 5, pp.505~515 (2005)
- Cox BD, "New concepts regarding focal adhesion kinase promotion of cell migration and proliferation", J Cell Biochem, 99, pp.35~52 (2006)
- Liu S, "FAK is required for TGFbeta-induced JNK phosphorylation in fibroblasts: implications for acquisition of a matrix-remodeling phenotype", Mol Biol Cell, 18, pp.2169~2178 (2007)
- Horowitz JC, "Combinatorial activation of FAK and AKT by transforming growth factor-beta1 confers an anoikis-resistant phenotype to myofibroblasts", Cell Signal, 19, pp.761~771 (2007)
- Parsons CJ, "Molecular mechanisms of hepatic fibrogenesis", J Gastroenterol Hepatol, 22(1), pp.79~84 (2007)
- Walsh MF, "Transforming growth factor-beta stimulates intestinal epithelial focal adhesion kinase synthesis via Smad- and p38-dependent mechanisms", Am J Pathol, 173, pp.385~399 (2008)
- Walsh MF, "TGF-beta1 modulates focal adhesion kinase expression in rat intestinal epithelial IEC-6 cells via stimulatory and inhibitory Smad binding elements", Biochim Biophys Acta, 1789, pp.88~98 (2009)
- Cordeiro MF, "Technology evaluation: lerdelimumab, Cambridge Antibody Technology", Curr Opin Mol Ther, 5, pp.199~203 (2003)
- Mead AL, "Evaluation of anti-TGF-beta2 antibody as a new postoperative anti-scarring agent in glaucoma surgery", Invest Ophthalmol Vis Sci, 44, pp.3394~3401 (2003)
- CAT-152 0102 Trabeculectomy Study Group, "A phase III study of subconjunctival human anti-transforming growth factor beta(2) monoclonal antibody (CAT-152) to prevent scarring after first-time trabeculectomy", Ophthalmology, 114, pp.1822~1830 (2007)