Transforming Growth Factor-β Stimulates Articular Chondrocyte Cell Growth through p44/42 MAP Kinase (ERK) Activation

• Published in: Endocrine Journal 1999, Vol.46(4), pp.545-553
• Main Authors: YONEKURA, AKIHIKO, OSAKI, MAKOTO, HIROTA, YASUHIRO, TSUKAZAKI, TOMOO, MIYAZAKI, YOUICHI, MATSUMOTO, TOMOKO, OHTSURU, AKIRA, NAMBA, HIROYUKI, SHINDO, HIROYUKI, YAMASHITA, SHUNICHI
• Format: Article
• Language: English
• Subjects: Animals; Articular chondrocyte; Cell Division - drug effects; Cell growth; Chondrocytes - cytology; Chondrocytes - drug effects; Dimethyl Sulfoxide - pharmacology; Enzyme Activation; ERK; extracellular signal-regulated kinase; Male; MAPK; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases - metabolism; Phosphorylation; Rats; Rats, Sprague-Dawley; Solvents - pharmacology; TGF-β1; Transforming Growth Factor beta - pharmacology; Transforming growth factor-^b1
• ISSN: 0918-8959; 1348-4540
• DOI: 10.1507/endocrj.46.545

Transforming growth factor-β1 (TGF-β1) stimulates articular chondrocyte cell proliferation and extracellular matrix formation. We reported previously that immediate and transient expression of c-fos mRNA through protein kinase C activation is required for the mitogenic effect of TGF-β1 on cultured rat articular chondrocytes (CRAC). In gel kinase assays using myelin basic protein (MBP) showed that total cell lysates from cells treated with TGF-β1 caused rapid phosphorylation of MBP, which suggests the involvement of mitogen-activated protein kinase (MAPK) activation. To identify specific MAPK pathways activated by TGF-β1, we performed in vitro kinase assays using specific substrates. TGF-β1 induced a rapid activation of extracellular signal regulated kinase (ERK) with a peak at 5min, which decreased to basal levels within 240min after TGF-β1 stimulation. In contrast, the c-jun N-terminal kinase activity increased only about 2.5-fold after 240min of stimulation and p38 MAPK activity did not change significantly. ERK activation by TGF-β1 was also confirmed by in vivo phosphorylation assays of Elk1. However, a specific MEK1 inhibitor, PD98059, significantly decreased TGF-β1 induced Elk1 phosphorylation in a dose-dependent manner. Furthermore, PD98059 reduced the TGF-β1-induced cell growth by 40%. These results indicate that TGF-β1 specifically activates MEK1 and subsequent ERK pathways in CRAC, and that the activation of this MAPK pathway plays a role in the mitogenic response to TGF-β1.