Abstract
Mucus hypersecretion is a common pathological change in chronic inflammatory diseases of the airway. These conditions are usually accompanied by chronic mechanical stress due to airway constriction. Our objective was to study the molecular mechanisms and physical effects of chronic mechanical stress on mucin 5AC (MUC5AC) expression in airway epithelial cells. We exposed normal human bronchial epithelial (NHBE) cells cultured at an air–liquid interface to different degrees of chronic compressive mechanical stress (10, 20, 30 cmH2O) for 7 days(1 h per day). MUC5AC protein content was detected by enzyme-linked immunosorbent assay (ELISA). MUC5AC mRNA expression was detected by reverse transcription PCR (RT-PCR) and real-time PCR. The effects of chronic mechanical stress on phosphorylated ERK1/2 (p-ERK1/2), phosphorylated JNK (p-JNK), phosphorylated P38 (p-P38), and phosphorylation of FAK at Tyr397 (p-FAK-Y397), were assessed by Western blot. We also assessed the impact of, an EGFR kinase inhibitor (AG1478), an ERK kinase inhibitor (PD-98059), and short interfering RNA (siRNA) targeted to FAK. We found that transcriptional and protein expression levels of MUC5AC were elevated significantly in the 30 cmH2O compressive stress group. p-ERK1/2 increased significantly in response to compressive stress and PD-98059 could attenuated stress-induced MUC5AC expression. p-FAK-Y397 increased significantly in response to compressive stress and FAK siRNA attenuated stress-induced ERK activation strongly. AG1478 attenuated stress-induced ERK activation and MUC5AC expression significantly, but incompletely. Combination of FAK siRNA and AG1478 led to complete attenuation of ERK activation and MUC5AC expression. These results suggest that chronic mechanical stress can enhance MUC5AC expression in human bronchial epithelial cells through the ERK signal transduction pathway. Both FAK and EGFR mediate the mitogenic response induced by mechanical stress in human bronchial epithelial cells through an ERK signaling cascade.
Similar content being viewed by others
References
Fahy JV (2001) Remodeling of the airway epithelium in asthma. Am J Respir Crit Care Med 164:S46–S51
Hogg JC, Chu F, Utokaparch S, Woods R, Elliott WM, Buzatu L, Cherniack RM, Rogers RM, Sciurba FC, Coxson HO, Paré PD (2004) The nature of small-airway obstruction in chronic obstructive pulmonary disease. N Engl J Med 350:2645–2653
Ha U, Lim JH, Jono H, Koga T, Srivastava A, Malley R, Pagès G, Pouysségur J, Li JD (2007) A novel role for IkappaB kinase (IKK) alpha and IKKbeta in ERK-dependent up-regulation of MUC5AC mucin transcription by Streptococcus pneumoniae. J Immunol 178:1736–1747
Qi Li, Zhou Xiang Dong, Xu Xiao Yan, Yang Jie (2010) Recombinant human elafin protects airway epithelium integrity during inflammation. Mol Biol Rep 37:2981–2988
Kim CH, Kim KE, Yoon JH, Song KS (2009) Upregulation of MUC5AC gene expression by IL-4 through CREB in human airway epithelial cells. J Cell Biochem 108:974–981
Zhou Xiang-Dong, Tong Jin, Lan Jian (2002) Chronic obstructive pulmonary disease airway mucin molecular phenotypic studies. Chin J Tuberc Respir Dis 25:437
Wiggs BR, Hrousis CA, Drazen JM, Kamm RD (1997) On the mechanism of mucosal folding in normal and asthmatic airways. J Appl Physiol 83:1814–1821
Tschumperlin DJ, Shively JD, Swartz MA, Silverman ES, Haley KJ, Raab G, Drazen JM (2002) Bronchial epithelial compression regulates MAP kinase signaling and HB-EGF-like growth factor expression. Am J Physiol Lung Cell Mol Physiol 282:L904–L911
Hewson CA, Edbrooke MR, Johnston SL (2003) PMA induces the MUC5AC respiratory mucin in human bronchial epithelial cells, via PKC, EGF/TGF-alpha, Ras/Raf, MEK, ERK and Sp1-dependent mechanisms. J Mol Biol 344:683–695
Song KS, Lee WJ, Chung KC, Koo JS, Yang EJ, Choi JY, Yoon JH (2003) Interleukin-1 beta and tumor necrosis factor-alpha induce MUC5AC overexpression through a mechanism involving ERK/p38 mitogen-activated protein kinases-MSK1-CREB activation in human airway epithelial cells. J Biol Chem 278:23243–23250
Song JS, Cho KS, Yoon HK, Moon HS, Park SH (2005) Neutrophil elastase causes MUC5AC mucin synthesis via EGF receptor, ERK and NF-kB pathways in A549 cells. Korean J Intern Med 20:275–283
Chaturvedi LS, Marsh HM, Basson MD (2007) Src and focal adhesion kinase mediate mechanical strain-induced proliferation and ERK1/2 phosphorylation in human H441 pulmonary epithelial cells. Am J Phyiol Cell Physiol 292:C1701–C1713
Tschumperlin DJ, Dai G, Maly IV, Kikuchi T, Laiho LH, McVittie AK, Haley KJ, Lilly CM, So PT, Lauffenburger DA, Kamm RD, Drazen JM (2004) Mechanotransduction through growth-factor shedding into the extracellular space. Nature 429:83–86
Kikuchi T, Shively JD, Foley JS, Drazen JM, Tschumperlin DJ (2004) Differentiation-dependent responsiveness of bronchial epithelial cells to IL-4/13 stimulation. Am J Physiol Lung Cell Mol Physiol 287:L119–L126
Park JA, Tschumperlin DJ (2009) Chronic intermittent mechanical stress increases MUC5AC protein expression. Am J Respir Cell Mol Biol 41:459–466
Ross AJ, Dailey LA, Brighton LE, Devlin RB (2007) Transcriptional profiling of mucociliary differentiation in human airway epithelial cells. Am J Respir Cell Mol Biol 37:169–185
Ressler B, Lee RT, Randell SH, Drazen JM, Kamm RD (2000) Molecular responses of rat tracheal epithelial cells to transmembrane pressure. Am J Physiol Lung Cell Mol Physiol 278:L1264–L1272
Thamilselvan V, Basson MD (2004) Pressure activates colon cancer cell adhesion by inside-out focal adhesion complex and actin cytoskeletal signaling. Gastroenterology 126:8–18
Yuan-Chen Wu D, Wu R, Reddy SP, Lee YC, Chang MM (2007) Distinctive epidermal growth factor receptor/extracellular regulated kinase-independent and -dependent signaling pathways in the induction of airway mucin 5B and mucin 5AC expression by phorbol 12-myristate 13-acetate. Am J Pathol 170:20–32
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25:402–408
Takami K, Takuwa N, Okazaki H, Kobayashi M, Ohtoshi T, Kawasaki S, Dohi M, Yamamoto K, Nakamura T, Tanaka M, Nakahara K, Takuwa Y, Takizawa H (2002) Interferon-γ inhibits hepatocyte growth factor stimulated cell proliferation of human bronchial epithelial cells: upregulation of p27kip1 cyclin-dependent kinase inhibitor. Am J Respir Cell Mol Biol 26:231–238
Wirtz HR, Dobbs LG (2000) The effects of mechanical forces on lung functions. Respir Physiol 119:1–17
Liu M, Tanswell AK, Post M (1999) Mechanical force-induced signal transduction in lung cells. Am J Physiol 277:L667–L683
Liu M, Post M (2000) Mechanochemical signal transduction in the fetal lung. J Appl Physiol 89:2078–2084
Vlahakis NE, Hubmayr RD (2000) Plasma membrane stress failure in alveolar epithelial cells. J Appl Physiol 89:2490–2496
Rose MC, Voynow JA (2006) Respiratory tract mucin genes and mucin glycoproteins in health and disease. Physiol Rev 86:245–278
Zuhdi Alimam M, Piazza FM, Selby DM, Letwin N, Huang L, Rose MC (2000) Muc-5/5ac mRNA and protein expression is a marker of goblet cell metaplasia in murine airways. Am J Respir Cell Mol Biol 22:253–260
Chess PR, Toia L, Finkelstein JN (2000) Mechanical strain-induced proliferation and signaling in pulmonary epithelial H441 cells. Am J Physiol Lung Cell Mol Physiol 279:L43–L51
Oudin S, Pugin J (2002) Role of MAP kinase activation in interleukin-8 production by human BEAS-2B bronchial epithelial cells submitted to cyclic stretch. Am J Respir Cell Mol Biol 27:107–114
Kippenberger S, Bernd A, Loitsch S, Guschel M, Muller J, Bereiter-Hahn J, Kaufmann R (2000) Signaling of mechanical stretch in human keratinocytes via MAP kinases. J Invest Dermatol 114:408–412
Li W, Duzgun A, Sumpio BE, Basson MD (2001) Integrin and FAK-mediated MAPK activation is required for cyclic strain mitogenic effects in caco-2 cells. Am J Physiol Gastrointest Liver Physiol 280:G75–G87
Peverali FA, Basdra EK, Papavassiliou AG (2001) Stretch-mediated activation of selective MAPK subtypes and potentiation of AP-1 binding in human osteoblastic cells. Mol Med 7:68–78
Zhang J, Li W, Sanders MA, Sumpio BE, Panja A, Basson MD (2003) Regulation of the intestinal epithelial response to cyclic strain by extracellular matrix proteins. FASEB J 17:926–928
Chess PR, O’Reilly MA, Sachs F, Finkelstein JN (2005) Reactive oxidant and p42/44 MAP kinase signaling is necessary for mechanical strain-induced proliferation in pulmonary epithelial cells. J Appl Physiol 99:1226–1232
Alexander LD, Alagarsamy S, Douglas JG (2004) Cyclic stretch-induced cPLA2 mediates ERK 1/2 signaling in rabbit proximal tubule cells. Kidney Int 65:551–563
Eric KC, John SF, Cheng Jason, Anita SP, Jeffrey MD, Tschumperlin DJ (2005) Bronchial epithelial compression regulates epidermal growth factor receptor family ligand expression in an autocrine manner. Am J Respir Cell Mol Biol 32:373–380
Acknowledgments
This work was supported by a grant from the National Nature Science Foundation of China (No.81070031), and the China-Russia Cooperation Research Foundation (No.81011120108).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, N., Li, Q., Zhou, X.D. et al. Chronic mechanical stress induces mucin5AC expression in human bronchial epithelial cells through ERK dependent pathways. Mol Biol Rep 39, 1019–1028 (2012). https://doi.org/10.1007/s11033-011-0827-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-011-0827-x