This Article Full Text Full Text (PDF) Data Supplement All Versions of this Article: 25/2/341    most recent 01.ATV.0000152608.29351.8fv1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sato, M. Articles by Kurabayashi, M. Search for Related Content PubMed PubMed Citation Articles by Sato, M. Articles by Kurabayashi, M. Pubmed/NCBI databases Gene GEO Profiles HomoloGene UniGene Compound via MeSH Substance via MeSH Hazardous Substances DB ACETYLCYSTEINE HYDROGEN PEROXIDE SODIUM DIETHYLDITHIOCARBAMATE Related Collections Oxidant stress Cell signalling/signal transduction Gene expression Hypertension - basic studies

(Arteriosclerosis, Thrombosis, and Vascular Biology. 2005;25:341.)
© 2005 American Heart Association, Inc.

Vascular Biology

c-Src and Hydrogen Peroxide Mediate Transforming Growth Factor-ß1–Induced Smooth Muscle Cell–Gene Expression in 10T1/2 Cells

Mahito Sato; Keiko Kawai-Kowase; Hiroko Sato; Yuko Oyama; Hiroyoshi Kanai; Yoshio Ohyama; Tatsuo Suga; Toshitaka Maeno; Yasuhiro Aoki; Junichi Tamura; Hironosuke Sakamoto; Ryozo Nagai; Masahiko Kurabayashi

From the Departments of Medicine and Biological Science (M.S., K.K.-K., H.S., Y. Oyama, H.K., Y. Ohyama, T.S., T.M., Y.A., M.K.) and General Medicine (M.S., J.T., H.S.), Gunma University Graduate School of Medicine, Japan; and the Department of Cardiovascular Medicine (R.N.), Graduate School of Medicine, University of Tokyo, Japan.

Correspondence to Masahiko Kurabayashi, MD, PhD, Department of Medicine and Biological Science, Gunma University Graduate School of Medicine, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan. E-mail mkuraba{at}med.gunma-u.ac.jp

Objective— Transforming growth factor-ß1 (TGF-ß1) controls the expression of numerous genes, including smooth muscle cell (SMC)–specific genes and extracellular matrix protein genes. Here we investigated whether c-Src plays a role in TGF-ß1 signaling in mouse embryonic fibroblast C3H10T1/2 cells.

Methods and Results— TGF-ß1 induction of the SMC contractile protein SM22 gene expression was inhibited by PP1 (an inhibitor of Src family kinases) or by C-terminal Src kinase (a negative regulator of c-Src). Induction of SM22 by TGF-ß1 was markedly attenuated in SYF cells (c-Src^–, Yes^–, and Fyn^–) compared with Src⁺⁺ cells (c-Src⁺⁺, Yes^–, and Fyn^–). PP1 also inhibited the TGF-ß1–induced expression of serum response factor (SRF), a transcription factor regulating the SMC marker gene expression. Confocal immunofluorescence analysis showed that TGF-ß1 stimulates production of hydrogen peroxide. Antioxidants such as catalase or NAD(P)H oxidase inhibitors such as apocynin inhibited the TGF-ß1–induced expression of SM22. Furthermore, we demonstrate that TGF-ß1 induction of the plasminogen activator inhibitor-1 (PAI-1) gene, which is known to be dependent on Smad but not on SRF, is inhibited by PP1 and apocynin.

Conclusion— Our results suggest that TGF-ß1 activates c-Src and generates hydrogen peroxide through NAD(P)H oxidase, and these signaling pathways lead to the activation of specific sets of genes, including SM22 and PAI-1.

TGF-ß1 controls the expression of numerous genes, including SM22 and PAI-1. We investigated whether c-Src plays a role in TGF-ß1 signaling. TGF-ß1 induction of such genes was significantly reduced in Src family tyrosine kinase–deficient cells, and Csk and pharmacological inhibitors for Src family kinases or antioxidants inhibit the effects of TGF-ß1. These results indicate that c-Src and hydrogen peroxide are required for TGF-ß1 signaling.

Key Words: growth factors • cytokines • oxidant stress • gene expression • smooth muscle cells

This article has been cited by other articles:

				F. Peng, B. Zhang, D. Wu, A. J. Ingram, B. Gao, and J. C. Krepinsky TGF{beta}-induced RhoA activation and fibronectin production in mesangial cells require caveolae Am J Physiol Renal Physiol, July 1, 2008; 295(1): F153 - F164. [Abstract] [Full Text] [PDF]

				C. H. Byon, A. Javed, Q. Dai, J. C. Kappes, T. L. Clemens, V. M. Darley-Usmar, J. M. McDonald, and Y. Chen Oxidative Stress Induces Vascular Calcification through Modulation of the Osteogenic Transcription Factor Runx2 by AKT Signaling J. Biol. Chem., May 30, 2008; 283(22): 15319 - 15327. [Abstract] [Full Text] [PDF]

				J. N Artaza, R. Singh, M. G Ferrini, M. Braga, J. Tsao, and N. F Gonzalez-Cadavid Myostatin promotes a fibrotic phenotypic switch in multipotent C3H 10T1/2 cells without affecting their differentiation into myofibroblasts J. Endocrinol., February 1, 2008; 196(2): 235 - 249. [Abstract] [Full Text] [PDF]

				A. K. Khanna and G. M. Pieper NADPH oxidase subunits (NOX-1, p22phox, Rac-1) and tacrolimus-induced nephrotoxicity in a rat renal transplant model Nephrol. Dial. Transplant., February 1, 2007; 22(2): 376 - 385. [Abstract] [Full Text] [PDF]

				M. Maeda, Y. Shintani, M. J. Wheelock, and K. R. Johnson Src Activation Is Not Necessary for Transforming Growth Factor (TGF)-{beta}-mediated Epithelial to Mesenchymal Transitions (EMT) in Mammary Epithelial Cells: PP1 DIRECTLY INHIBITS TGF-{beta} RECEPTORS I AND II J. Biol. Chem., January 6, 2006; 281(1): 59 - 68. [Abstract] [Full Text] [PDF]