Transcriptional activation of endothelial cell integrin alpha v by protein kinase C activator 12(S)-HETE
| Autor: | Clement A. Diglio, Kenneth V. Honn, Rajesh Bazaz, Dean G. Tang |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Transcriptional Activation
Pulmonary Circulation Alpha-v beta-3 Molecular Sequence Data Integrin Fluorescent Antibody Technique Biology Polymerase Chain Reaction Antibodies Mice chemistry.chemical_compound Antigens CD Hydroxyeicosatetraenoic Acids Gene expression Animals 12-Hydroxy-5 8 10 14-eicosatetraenoic Acid RNA Messenger Protein Kinase C Protein kinase C DNA Primers Cell Nucleus Base Sequence Activator (genetics) Cell Biology Integrin alphaV Blotting Northern Flow Cytometry Molecular biology Clone Cells Blotting Southern Calphostin C chemistry biology.protein Tetradecanoylphorbol Acetate Solution hybridization Endothelium Vascular |
| Zdroj: | Journal of Cell Science. 108:2629-2644 |
| ISSN: | 1477-9137 0021-9533 |
| DOI: | 10.1242/jcs.108.7.2629 |
| Popis: | Previous work demonstrated that 12(S)-HETE [12(S)-hydroxyeicosatetraenic acid], a lipoxygenase metabolite of arachidonic acid, stimulates the surface expression of integrin alpha v beta 3 on mouse lung vascular endothelial cells (CD clone 3) in a post-transcriptional and protein kinase C (PKC)-dependent fashion. In this study we examined the effect of 12(S)-HETE on the expression of integrin receptors alpha v beta 3 and alpha 5 beta 1 in a different clone of a mouse endothelial cell population derived from lung microvasculature (designated CD clone 4). The results indicated that 12(S)-HETE transcriptionally activates the gene expression of integrin alpha v as assessed by quantitative reverse transcription/polymerase chain reaction/Southern hybridization, RNase protection assay, solution hybridization, and northern blotting. The induction of alpha v mRNA occurred within 1 hour, peaked at approximately 4 hours (2- to 4-fold increase), persisted for up to 16 hours, and thereafter gradually declined. The PKC activator phorbol 12-myristate 13-acetate (PMA) induced the alpha v mRNA, in a similar way. 12(S)-HETE treatment did not, in contrast, alter the mRNA levels of integrin subunit alpha 5 or beta 1. The induction of alpha v mRNA appeared to be protein synthesis-independent, since cycloheximide did not alter the 12(S)-HETE effect. 12(S)-HETE also did not appear to alter the mRNA half-life of alpha v. On the other hand, 12(S)-HETE-induced increase in alpha v mRNA levels was PKC-dependent, since pretreatment of CD clone 4 cells with calphostin C significantly inhibited 12(S)-HETE-increased alpha v mRNA. Nuclear runoff experiments revealed that the increase in alpha v mRNA results from an enhanced gene transcription. Facilitated alpha v gene transcription resulted in an increased surface expression of alpha v beta 3 protein, which resulted in an increased cell adhesion to vitronectin. The above observations, in conjunction with our previous experimental data, suggest that 12(S)-HETE may employ diverse mechanisms to stimulate the integrin alpha v beta 3 expression in vascular endothelial cells, which could play important roles in tumor cell adhesion, angiogenesis, hemostasis, and many other vascular events. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|