RETRACTED ARTICLE: Thrombin stimulates VSMC proliferation through an EGFR-dependent pathway: involvement of MMP-2

Abstract

In this study, the role of epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase (ERK1/2), heparin-binding EGF-like growth factor (HB-EGF), general metalloproteinases, matrix metalloproteinases-2 (MMP-2) in mediating the mitogenic action of thrombin in rat vascular smooth muscle cells (VSMC) was investigated. The incubation of rat VSMC with thrombin (1 U/ml) for 5 min resulted in significant (p < 0.001) increase of ERK1/2 phosphorylation by 8.7 ± 0.9-fold, EGFR phosphorylation by 8.5 ± 1.3-fold (p < 0.001) and DNA synthesis by 3.6 ± 0.4-fold (p < 0.001). Separate 30-min pretreatments with EGFR tyrosine kinase irreversible inhibitor, 10 µM PD169540 (PD), and 20 µM anti-HB-EGF antibody significantly reduced thrombin-stimulated EGFR and ERK1/2 phosphorylation by 81, 72 % and by 48 and 61 %, respectively. Furthermore, the same pretreatments with PD or anti-HB-EGF antibody reduced thrombin-induced VSMC proliferation by 44 and 45 %, respectively. In addition, 30-min pretreatments with 10 µM specific MMP-2 inhibitor significantly reduced thrombin-stimulated phosphorylation of both EGFR and ERK1/2 by 25 %. Moreover, the same pretreatment with MMP-2 inhibitor reduced thrombin-induced VSMC proliferation by 45 %. These results show that the thrombin-induced DNA synthesis correlates with the level of ERK1/2 activation rather than EGFR activation. These results further suggest that thrombin acts through EGFR and ERK 1/2 signaling pathways involving MMP-2 to upregulate proliferation of VSMC.

Salusin-β contributes to vascular remodeling associated with hypertension via promoting vascular smooth muscle cell proliferation and vascular fibrosis

“…Heparinbinding EGF-like growth factor (HB-EGF) is recognized as a ligand that binds to EGFR [31]. HB-EGF is highly expressed in VSMCs and has been shown to stimulate proliferation of VSMCs via autophosphorylation of EGFR [32]. We found that anti-HB-EGF IgG prevented the salusin-β-induced VSMCs proliferation, attenuated the salusin-β-induced EGFR phosphorylation, and abolished the salusin-β-induced phosphorylation of CREB and ERK.…”

RNA sequencing to determine the contribution of kinase receptor transactivation to G protein coupled receptor signalling in vascular smooth muscle cells

“…Over the past two decades there have been studies expanding on GPCR mediated transactivation signalling of PTKR specifically the EGFR [39, 50, 51]. GPCR mediated transactivation dependent signalling also spans out to include activation of the TGFBR1.…”

Inhibitory effects of fermented extract of Ophiopogon japonicas on thrombin-induced vascular smooth muscle cells

“…These results suggested that FEOJ blocks FEOJ blocks thrombin-induced migration of VSMCs. Migration of VSMCs is a main step in atherosclerosis (1,2) and thrombin has been reported to induce migration of VSMCs (4,5). To investigate whether FEOJ inhibits thrombin-induced migration of VSMCs, a wound-healing assay was used.…”

“…After stimulation with thrombin, VSMCs undergo G1-to-S-phase cell-cycle transition, which is tightly regulated by cyclin D1 and p27KIP1 (2,3). In addition, matrix metalloproteinase-2 (MMP-2) degrades type IV collagen, leading to VSMC migration and the progression of arterial lesions (4,5). A previous study has demonstrated that thrombin stimulates the MMP-2 expression via transcription factor nuclear factor (NF)-κB in chondrosarcoma cells (6).…”