Src and Cas Are Essentially but Differentially Involved in Angiotensin II-Stimulated Migration of Vascular Smooth Muscle Cells via Extracellular Signal-Regulated Kinase 1/2 and c-Jun NH₂-Terminal Kinase Activation

Abstract

Angiotensin II (Ang II) plays an important role in several cardiovascular diseases associated with vascular smooth muscle cell (VSMC) growth and migration. Src activity is known to be required for the migration of a number of cell types. p130Cas was reported to be essential for cell migration and actin filament reorganization. Mitogen-activated protein (MAP) kinases were also reported to be critical regulatory factors for growth and migration of VSMC. However, precise intracellular mechanisms involving c-Src, p130Cas, and MAP kinases in Ang IIstimulated migration of VSMC have not been well elucidated. Here we demonstrated that Ang II rapidly and significantly stimulated tyrosine phosphorylation of Src and Cas and their association in rat aortic smooth muscle cells (RASMC). Ang II-stimulated tyrosine phosphorylation of Src and Cas and activation of ERK1/2 and JNK, but not p38, were potently inhibited by Src family tyrosine kinase inhibitors, herbimycin A (HA) and PP2. Ang II-stimulated Src and Cas association, tyrosine phosphorylation of Cas, and activation of ERK1/2 and JNK were suppressed in kinase-inactive Src (KI Src)-overexpressed RASMC. Ang II-stimulated JNK activation but not ERK1/2 activation was blocked in substrate domain-deleted Cas (⌬SD Cas)-overexpressed RASMC. In addition, HA, PP2, ERK1/2 inhibitor, 2Ј-amino-3Ј-methoxyflavone (PD98059) and JNK inhibitor, and anthra[1,9-cd]pyrazol-6(2H)-one (SP600125) significantly inhibited Ang II-stimulated migration of RASMC. Ang II-induced colocalization of Src and Cas and migration were inhibited in both KI Src-and ⌬SD Cas-overexpressed RASMC. These findings suggest that Src and Cas are essentially but differentially involved in Ang II-stimulated migration of VSMC through the activation of ERK1/2 and JNK.Angiotensin II (Ang II) plays an important role in several cardiovascular diseases associated with vascular smooth muscle cell (VSMC) growth (Berk et al., 1989) and migration (Griendling et al., 1996), such as atherosclerosis, restenosis after angioplasty, and hypertension. It was indicated that tyrosine phosphorylation is a critical episode in several Ang II-stimulated signaling events (Marrero et al., 1996). Ang II induces tyrosine phosphorylation of proteins with apparent molecular masses of 42, 44, 70 to 80, 110 to 130, and 190 kDa in VSMC (Tsuda et al., 1991). The 130-, 60-, 44-, and 42-kDa proteins that were tyrosine-phosphorylated in response to Ang II in VSMC were identified as p130Cas [a CT-10 -regulated kinase (Crk)-associated substrate] (Takahashi et al., 1998), 60-kDa c-Src, a nonreceptor tyrosine kinase (Ishida et al., 1995), and mitogen-activated protein (MAP) kinase isozymes (Tsuda et al., 1992), respectively.Cell migration requires cytoskeletal reorganization involving phosphorylation of cytoskeleton-associated tyrosine kinases and formation of focal-adhesion complexes (Ilic et al.,

Nifedipine Interferes with Migration of Vascular Smooth Muscle Cells via Inhibition of Pyk2-Src Axis

“…Nifedipine potently inhibited PDGF-induced Src activation and tyrosine phosphorylation of Cas, paxillin, and cortactin, all of which have been implicated in cell migration [16][17][18] . Indeed, Kyaw and colleagues reported that Cas is involved in Anginduced VSMC migration 23) , and we previously used RNA interference techniques to demonstrate that Cas, cortactin, and paxillin are required for VSMC migration (unpublished data). Thus, the present data suggest that nifedipine inhibits VSMC migration via the inhibition of Src activation.…”

p38 Mitogen-Activated Protein Kinase Contributes to Angiotensin II-Stimulated Migration of Rat Aortic Smooth Muscle Cells

“…MAPKs are believed to be associated with the migration and proliferation of vascular smooth muscle cells (15,16). Migration, proliferation, and MAPK activation can occur in vascular smooth muscle cells stimulated by Ang II (5,20,21). Growth factors and cytokines activate the p38 MAPK pathway, which mediates cell migration in tracheal smooth muscle cells (22).…”

“…The activation of p38 MAPK leads to the contraction, migration, and proliferation of vascular smooth muscle cells (17 -19). Ang II is known to stimulate the activation of MAPKs, including extracellular signal-regulated kinase (ERK) 1 / 2, p38 MAPK, and stress-activated protein kinase / c-Jun Nterminal kinase in vascular smooth muscle cells (9,20,21). Previously, we have shown that the p38 MAPK pathway is involved in spleen tyrosine kinase (Syk)-induced migration in response to platelet-derived growth factor (PDGF) in vascular smooth muscle cells (3).…”

Signal-Crosstalk Between Rho/ROCK and c-Jun NH 2 -Terminal Kinase Mediates Migration of Vascular Smooth Muscle Cells Stimulated by Angiotensin II

“…7,15 Therefore, we have investigated the possible signaling cross-talk between Rho/ROCK activation and the MAPK family activation by Ang II. As shown in Figure 2A, Ang II-induced ERK and p38MAPK phosphorylations were not affected by dnRho.…”