The role of akt in induced apoptosis via interferon-gamma and angiotensin II stimulation after ras transfection in vascular smooth muscle cells

Angiotensin II (Ang II) is related with both signalization mechanisms concerning apoptosis and anti-apoptosis. It is also known that Ang II triggers its anti-apoptotic effects via Akt/protein kinase B (PKB) in vascular smooth muscle cells (VSMCs). In addition, there are many sub signalization pathways, such as IKK (NF-KappaB ? cell survival) and p70S6K (protein synthesis) which are being triggered over Akt and also it was shown that apoptosis is being induced via such mitochondrial proteins Bad, Bcl?2 by active phosphorylated Akt.This contradiction, which Akt is having a part in both apoptosis and cell survival, overviewed in the studies, makes us to predict a hypothesis that Akt can be a target molecule, which can take the cell to apoptosis even in Ras over-expressed media.In our study, as to emphasize that Akt is a junction molecule especially for apoptosis in VSMCs, we have tried to establish a model, after which the cell culture media induced by Ang II, Akt phosphorylation was measured as well as we did the same assays with the same parameters in VSMC culture after being induced by interferon-gamma that takes the cells to apoptosis. Besides, phosphorylated Akt levels were observed in such a media, which activated apoptotic signalization pathways via interferon-gamma in VSMC culture formed by the transfection of Ras plasmids. Both in Ras over-expressed and normal cells, as to observe interactions between phosphorylated Akt and the sub-effector molecules such as Bad and Bcl-XL, the immunprecipitation assays were carried out and by doing caspas-3 assays the level of apoptotic signals have been demonstrated in Ras over-expressed VSMC culture.Briefly, what we have showed is that the Akt/PKB is a junction molecule and especially plays an individual role on going apoptosis in VSMCs. On the other hand, decreased levels of Akt/PKB phosphorylation and Bad/Bcl-XL complex formation, emphasized the possible apoptotic mechanisms in Ras over-expressed media can easily be manipulated, but the resident fact is just the direct contrary.