PKC-δ/c-Src-mediated EGF receptor transactivation regulates thrombin-induced COX-2 expression and PGE2 production in rat vascular smooth muscle cells
| Autor: | Chuen-Mao Yang, Chi-Chin Sun, Hsi-Lung Hsieh, Tze-Shyuan Wang |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
MAPK/ERK pathway
Vascular smooth muscle Myocytes Smooth Muscle Proto-Oncogene Proteins pp60(c-src) Biology GTP-Binding Protein alpha Subunits Gi-Go Dinoprostone Gene Expression Regulation Enzymologic Muscle Smooth Vascular Small hairpin RNA Rats Sprague-Dawley chemistry.chemical_compound Transactivation Thrombin EGF receptor medicine Animals Extracellular Signal-Regulated MAP Kinases Molecular Biology Protein kinase C Cells Cultured ERK1/2 Receptor transactivation NF-kappa B Cell Biology COX-2 Molecular biology Matrix Metalloproteinases Rats ErbB Receptors Transcription Factor AP-1 Protein Kinase C-delta chemistry Cyclooxygenase 2 Vascular smooth muscle cell GTP-Binding Protein alpha Subunits Gq-G11 Rottlerin medicine.drug |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. (9):1563-1575 |
| ISSN: | 0167-4889 |
| DOI: | 10.1016/j.bbamcr.2008.03.016 |
| Popis: | The thrombin/proteinase-activated receptors (PARs) have been shown to regulate smooth muscle cell proliferation, migration, and vascular maturation. Thrombin up-regulates expression of several proteins including cyclooxygenase (COX)-2 in vascular smooth muscle cells (VSMCs) and contributes to vascular diseases. However, the mechanisms underlying thrombin-regulated COX-2 expression in VSMCs remain unclear. Western blotting, RT-PCR, and EIA kit analyses showed that thrombin induced the expression of COX-2 mRNA and protein and PGE(2) release in a time-dependent manner, which was attenuated by inhibitors of PKC (GF109203X and rottlerin), c-Src (PP1), EGF receptor (EGFR; AG1478) and MEK1/2 (U0126), or transfection with dominant negative mutants of PKC-delta, c-Src or extracellular regulated kinase (ERK) and ERK1 short hairpin RNA interference (shRNA). These results suggest that transactivation of EGFR participates in COX-2 expression induced by thrombin in VSMCs. Accordingly, thrombin stimulated phosphorylation of ERK1/2 which was attenuated by GF109203X, rottlerin, PP1, GM6001, CRM197, AG1478, or U0126, respectively. Furthermore, this up-regulation of COX-2 mRNA and protein was blocked by selective inhibitors of AP-1 and NF-kappaB, curcumin and helenalin, respectively. Moreover, thrombin-stimulated activation of NF-kappaB, AP-1, and COX-2 promoter activity was blocked by the inhibitors of c-Src, PKC, EGFR, MEK1/2, AP-1 and NF-kappaB, suggesting that thrombin induces COX-2 promoter activity mediated through PKC(delta)/c-Src-dependent EGFR transactivation, MEK-ERK1/2, AP-1, and NF-kappaB. These results demonstrate that in VSMCs, activation of ERK1/2, AP-1 and NF-kappaB pathways was essential for thrombin-induced COX-2 gene expression. Understanding the regulation of COX-2 expression and PGE(2) release by thrombin/PARs system on VSMCs may provide potential therapeutic targets of vascular inflammatory disorders including arteriosclerosis. |
| Databáze: | OpenAIRE |
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