Am. J. Respir. Cell Mol. Biol., Volume 21, Number 1, July, 1999 77-88

Glucocorticoids Inhibit Proliferation, Cyclin D1 Expression, and Retinoblastoma Protein Phosphorylation, but Not Activity of the Extracellular-Regulated Kinases in Human Cultured Airway Smooth Muscle

Darren Fernandes, Elizabeth Guida, Valentina Koutsoubos, Trudi Harris, Peter Vadiveloo, John W. Wilson, and Alastair G. Stewart

Department of Pharmacology, University at Melbourne; Bernard O'Brien Institute of Microsurgery, St. Vincent's Hospital, Melbourne; and Department of Respiratory Medicine, Alfred Hospital, Melbourne, Australia

We have previously shown that glucocorticoids inhibit mitogen-stimulated proliferation of human cultured airway smooth muscle (ASM) cells. The present study analyzed the effect of glucocorticoids on key regulatory pathways leading to passage of cells through the restriction point of the cell cycle, including those mediated by extracellular-regulated kinases (ERK) 1 and 2; the ERK upstream regulator MAPK kinase (MEK1); cyclin D1 levels; and levels and phosphorylation of retinoblastoma protein (pRb). Fluticasone propionate, a new inhaled glucocorticoid, was at least 10-fold more potent than dexamethasone in inhibiting thrombin-stimulated DNA synthesis and increases in cell number. Thrombin-stimulated increases in the levels and hyperphosphorylation of pRb were inhibited by glucocorticoids, which also reduced thrombin-stimulated cyclin D1 protein and messenger RNA (mRNA) levels. PD98059 (10 µM), an inhibitor of MEK1 activation, markedly attenuated thrombin stimulation of ERK activity and phosphorylation, DNA synthesis, and cyclin D1 levels. However, glucocorticoids had no effect on ERK activity or phosphorylation at 5 min, 2 h, or 12 h after addition of thrombin. In conclusion, glucocorticoid-induced reduction of cyclin D1 mRNA and protein levels, and of pRb phosphorylation, is sufficient to account for inhibition of ASM proliferation. Furthermore, these inhibitory effects of glucocorticoids on cyclin D1 and pRb occur on a component of the mitogen signaling cascade that is either downstream of or parallel to the ERK pathway.

This article has been cited by other articles:

				J. E. Ward, T. Harris, T. Bamford, A. Mast, M. C. F. Pain, C. Robertson, D. Smallwood, T. Tran, J. Wilson, and A. G. Stewart Proliferation is not increased in airway myofibroblasts isolated from asthmatics Eur. Respir. J., August 1, 2008; 32(2): 362 - 371. [Abstract] [Full Text] [PDF]

				M. Baroffio, E. Crimi, and V. Brusasco Review: Airway smooth muscle as a model for new investigative drugs in asthma Therapeutic Advances in Respiratory Disease, June 1, 2008; 2(3): 129 - 139. [Abstract] [PDF]

				J. K. Bentley and M. B. Hershenson Airway Smooth Muscle Growth in Asthma: Proliferation, Hypertrophy, and Migration Proceedings of the ATS, January 1, 2008; 5(1): 89 - 96. [Abstract] [Full Text] [PDF]

				M. C. Shepherd, S. M. Duffy, T. Harris, G. Cruse, M. Schuliga, C. E. Brightling, C. B. Neylon, P. Bradding, and A. G. Stewart KCa3.1 Ca2+Activated K+ Channels Regulate Human Airway Smooth Muscle Proliferation Am. J. Respir. Cell Mol. Biol., November 1, 2007; 37(5): 525 - 531. [Abstract] [Full Text] [PDF]

				I. S. T. Bos, R. Gosens, A. B. Zuidhof, D. Schaafsma, A. J. Halayko, H. Meurs, and J. Zaagsma Inhibition of allergen-induced airway remodelling by tiotropium and budesonide: a comparison Eur. Respir. J., October 1, 2007; 30(4): 653 - 661. [Abstract] [Full Text] [PDF]

				S. Xie, M. B. Sukkar, R. Issa, N. M. Khorasani, and K. F. Chung Mechanisms of induction of airway smooth muscle hyperplasia by transforming growth factor-beta Am J Physiol Lung Cell Mol Physiol, July 1, 2007; 293(1): L245 - L253. [Abstract] [Full Text] [PDF]

				S. S. An, T. R. Bai, J. H. T. Bates, J. L. Black, R. H. Brown, V. Brusasco, P. Chitano, L. Deng, M. Dowell, D. H. Eidelman, et al. Airway smooth muscle dynamics: a common pathway of airway obstruction in asthma Eur. Respir. J., May 1, 2007; 29(5): 834 - 860. [Abstract] [Full Text] [PDF]

				A. M. Goldsmith, M. B. Hershenson, M. P. Wolbert, and J. K. Bentley Regulation of airway smooth muscle {alpha}-actin expression by glucocorticoids Am J Physiol Lung Cell Mol Physiol, January 1, 2007; 292(1): L99 - L106. [Abstract] [Full Text] [PDF]

				I J Bujalska, M Quinkler, J W Tomlinson, C T Montague, D M Smith, and P M Stewart Expression profiling of 11{beta}-hydroxysteroid dehydrogenase type-1 and glucocorticoid-target genes in subcutaneous and omental human preadipocytes. J. Mol. Endocrinol., October 1, 2006; 37(2): 327 - 340. [Abstract] [Full Text] [PDF]

				S. Takayama, I. Rogatsky, L. E. Schwarcz, and B. D. Darimont The Glucocorticoid Receptor Represses Cyclin D1 by Targeting the Tcf-beta-Catenin Complex J. Biol. Chem., June 30, 2006; 281(26): 17856 - 17863. [Abstract] [Full Text] [PDF]

				B. Fouty, T. Moss, V. Solodushko, and M. Kraft Dexamethasone can stimulate G1-S phase transition in human airway fibroblasts in asthma Eur. Respir. J., June 1, 2006; 27(6): 1160 - 1167. [Abstract] [Full Text] [PDF]

				W. W. Woodmansee, J. M. Kerr, E. A. Tucker, J. R. Mitchell, D. J. Haakinson, D. F. Gordon, E. C. Ridgway, and W. M. Wood The Proliferative Status of Thyrotropes Is Dependent on Modulation of Specific Cell Cycle Regulators by Thyroid Hormone Endocrinology, January 1, 2006; 147(1): 272 - 282. [Abstract] [Full Text] [PDF]

				R. A. Panettieri Jr. Effects of Corticosteroids on Structural Cells in Asthma and Chronic Obstructive Pulmonary Disease Proceedings of the ATS, November 1, 2004; 1(3): 231 - 234. [Abstract] [Full Text] [PDF]

				C. Stellato Post-transcriptional and Nongenomic Effects of Glucocorticoids Proceedings of the ATS, November 1, 2004; 1(3): 255 - 263. [Abstract] [Full Text] [PDF]

				M. Roth, P. R.A. Johnson, P. Borger, M. P. Bihl, J. J. Rudiger, G. G. King, Q. Ge, K. Hostettler, J. K. Burgess, J. L. Black, et al. Dysfunctional Interaction of C/EBP{alpha} and the Glucocorticoid Receptor in Asthmatic Bronchial Smooth-Muscle Cells N. Engl. J. Med., August 5, 2004; 351(6): 560 - 574. [Abstract] [Full Text] [PDF]

				J. Tamaoki, E. Tagaya, K. Kawatani, J. Nakata, Y. Endo, and A. Nagai Airway Mucosal Thickening and Bronchial Hyperresponsiveness Induced by Inhaled {beta}2-Agonist in Mice Chest, July 1, 2004; 126(1): 205 - 212. [Abstract] [Full Text] [PDF]

				E. A. Goncharova, C. K. Billington, C. Irani, A. V. Vorotnikov, V. A. Tkachuk, R. B. Penn, V. P. Krymskaya, and R. A. Panettieri Jr. Cyclic AMP-Mobilizing Agents and Glucocorticoids Modulate Human Smooth Muscle Cell Migration Am. J. Respir. Cell Mol. Biol., July 1, 2003; 29(1): 19 - 27. [Abstract] [Full Text] [PDF]

				Y. Amrani, O. Tliba, D. Choubey, C.-D. Huang, V. P. Krymskaya, A. Eszterhas, A. L. Lazaar, and R. A. Panettieri Jr. IFN-gamma inhibits human airway smooth muscle cell proliferation by modulating the E2F-1/Rb pathway Am J Physiol Lung Cell Mol Physiol, June 1, 2003; 284(6): L1063 - L1071. [Abstract] [Full Text] [PDF]

				P A Beckett and P H Howarth Pharmacotherapy and airway remodelling in asthma? Thorax, February 1, 2003; 58(2): 163 - 174. [Abstract] [Full Text] [PDF]

				Y. Fujitani and A. Trifilieff In Vivo and In Vitro Effects of SAR 943, a Rapamycin Analogue, on Airway Inflammation and Remodeling Am. J. Respir. Crit. Care Med., January 15, 2003; 167(2): 193 - 198. [Abstract] [Full Text] [PDF]

				A. K. Greenberg, J. Hu, S. Basu, J. Hay, J. Reibman, T.-a. Yie, K. M. Tchou-Wong, W. N. Rom, and T. C. Lee Glucocorticoids Inhibit Lung Cancer Cell Growth through Both the Extracellular Signal-Related Kinase Pathway and Cell Cycle Regulators Am. J. Respir. Cell Mol. Biol., September 1, 2002; 27(3): 320 - 328. [Abstract] [Full Text] [PDF]

				K. Page, J. Li, K. C. Corbit, K. M. Rumilla, J.-W. Soh, I. B. Weinstein, C. Albanese, R. G. Pestell, M. R. Rosner, and M. B. Hershenson Regulation of Airway Smooth Muscle Cyclin D1 Transcription by Protein Kinase C-{delta} Am. J. Respir. Cell Mol. Biol., August 1, 2002; 27(2): 204 - 213. [Abstract] [Full Text] [PDF]

				R. A. Hughes, T. Harris, E. Altmann, D. McAllister, R. Vlahos, A. Robertson, M. Cushman, Z. Wang, and A. G. Stewart 2-Methoxyestradiol and Analogs as Novel Antiproliferative Agents: Analysis of Three-Dimensional Quantitative Structure-Activity Relationships for DNA Synthesis Inhibition and Estrogen Receptor Binding Mol. Pharmacol., May 1, 2002; 61(5): 1053 - 1069. [Abstract] [Full Text] [PDF]

				H. Witschi, I. Espiritu, M. Suffia, and K. E. Pinkerton Expression of cyclin D1/2 in the lungs of strain A/J mice fed chemopreventive agents Carcinogenesis, February 1, 2002; 23(2): 289 - 294. [Abstract] [Full Text] [PDF]

				A. M. Freyer, S. R. Johnson, and I. P. Hall Effects of Growth Factors and Extracellular Matrix on Survival of Human Airway Smooth Muscle Cells Am. J. Respir. Cell Mol. Biol., November 1, 2001; 25(5): 569 - 576. [Abstract] [Full Text] [PDF]

				A. J. Ammit and R. A. Panettieri Jr. Signal Transduction in Smooth Muscle: Invited Review: The circle of life: cell cycle regulation in airway smooth muscle J Appl Physiol, September 1, 2001; 91(3): 1431 - 1437. [Abstract] [Full Text] [PDF]

				T. E. Konopka, J. E. Barker, T. L. Bamford, E. Guida, R. L. Anderson, and A. G. Stewart Nitric Oxide Synthase II Gene Disruption: Implications for Tumor Growth and Vascular Endothelial Growth Factor Production Cancer Res., April 1, 2001; 61(7): 3182 - 3187. [Abstract] [Full Text]

				P. R. A. JOHNSON, J. L. BLACK, S. CARLIN, Q. GE, and P. ANNE UNDERWOOD The Production of Extracellular Matrix Proteins by Human Passively Sensitized Airway Smooth-Muscle Cells in Culture . The Effect of Beclomethasone Am. J. Respir. Crit. Care Med., December 1, 2000; 162(6): 2145 - 2151. [Abstract] [Full Text]

				A. G. Stewart, T. Harris, D. J. Fernandes, L. C. Schachte, V. Koutsoubos, E. Guida, C. E. Ravenhall, P. Vadiveloo, and J. W. Wilson beta 2-Adrenergic Receptor Agonists and cAMP Arrest Human Cultured Airway Smooth Muscle Cells in the G1 Phase of the Cell Cycle: Role of Proteasome Degradation of Cyclin D1 Mol. Pharmacol., November 1, 1999; 56(5): 1079 - 1086. [Abstract] [Full Text]