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ANTIAPOPTOTIC SIGNALING BY TF/FVIIA IS INHIBITED BY SIMVASTATIN
Abstract number: P-S-001
Åberg1 M., Wickström1 M., Siegbahn1 A.
1Medical Sciences, Uppsala University, Uppsala, Sweden
How-to-cite Åberg M, Wickström M, Siegbahn A. ANTIAPOPTOTIC SIGNALING BY TF/FVIIA IS INHIBITED BY SIMVASTATIN. J Thromb Haemost 2007; 5 Supplement 2: P-S-001
Abstract
Introduction: TF/FVIIa has been shown to induce antiapoptosis during serum starvation in a PI3-kinase/AKT dependent manner. Data on TF/FVIIa-mediated effects on apoptosis induced by other mechanisms are, however, lacking. We analyzed the influence of TF/FVIIa on simvastatin-induced apoptosis in human breast cancer cells.
Methods: MDA-MB-231 cells were incubated with or without 5 mM simvastatin and 10 or 100 nM FVIIa. Caspase-3, nuclear fragmentation, and NFkB translocation were measured using the ArrayScan HCS reader, an automated fluorescence imaging microscope. mRNA for b-actin, BCL-2, and AKT1 were analyzed with RT-PCR while protein levels and phosphorylation of AKT were determined by western blotting.
Results: Simvastatin induced apoptosis after 48 h of treatment. The levels of caspase-3 and nuclear fragmentation was by then increased four and two times respectively as compared to untreated controls. Incubation with 100 nM FVIIa resulted in a twofold increase of AKT phosphorylation at 24 h. However, in the presence of 5 mM simvastatin we found no AKT phosphorylation or anti-apoptotic effect, assessed as caspase-3, of the TF/FVIIa signaling complex. After 12 h of incubation with 5 mM simvastatin, analyses of the spatial translocation of NFkB showed 50% retention of NFkB in the cytoplasm. A number of genes were affected by this and the mRNA expression of the antiapoptotic protein BCL2 was diminished with 50% at 12 h and not detectable at 24 h. AKT1 mRNA was still unaffected at 12 h but reduced with 75% at 24 h.
Conclusions: The proapoptotic effect of simvastatin was due to a transcriptional down regulation of the antiapoptotic protein BCL-2. Since the PI3-kinase/AKT signaling pathway was quenched, FVIIa was unable to inhibit the simvastatin induced apoptosis.
To cite this abstract use the following format:
Journal of Thrombosis and Haemostasis 2007; Volume 5, Supplement 2: abstract number
Session Details
| Date: |
01/08/2007
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| Time: |
00:00-00:00
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| Session name: |
XXIst ISTH Congress |
| Subject: |
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| Location: |
Oxford, UK |
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