The experimental study on phosphatidylinositol 3-kinase/Akt in neural stem cells in vitro
Shuang Wang
Abstract
Shuang Wang
Abstract
Objective:To discuss the function of PI3K (phosphatidylinositol-3 kinase)/Akt signal pathway in proliferation and differentiation of neural stem cells (NSCs).Methods:Control group,wortmannin group and LY294002 group were randomly assigned.The expression of Nestin,NSE and S100 were detected by immunohistochemistry.The ratio of cell apoptosis was estimated by TUNEL staining and the expressions of phosphorylated Akt and Caspase-9 were detected by Western blotting.Results:With the increase in concentration of wortmannin and LY294002,the ratio of NSCs apoptosis increased.The results of Western blotting indicated that the expression of phosphorylated Akt decreased when the concentration of wortmannin and LY294002 increased.High concentration of wortmannin and LY294002 inhitited Akt phosphorylated.At the same time,the expression of phosphorylated Caspase-9 decreased.Conclusion:The survival of NSCs depends on the activation of PI3K/Akt pathway,but PI3K/Akt pathway may have little function in the differentiation of NSCs.
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Objective:To discuss the function of PI3K (phosphatidylinositol-3 kinase)/Akt signal pathway in proliferation and differentiation of neural stem cells (NSCs).Methods:Control group,wortmannin group and LY294002 group were randomly assigned.The expression of Nestin,NSE and S100 were detected by immunohistochemistry.The ratio of cell apoptosis was estimated by TUNEL staining and the expressions of phosphorylated Akt and Caspase-9 were detected by Western blotting.Results:With the increase in concentration of wortmannin and LY294002,the ratio of NSCs apoptosis increased.The results of Western blotting indicated that the expression of phosphorylated Akt decreased when the concentration of wortmannin and LY294002 increased.High concentration of wortmannin and LY294002 inhitited Akt phosphorylated.At the same time,the expression of phosphorylated Caspase-9 decreased.Conclusion:The survival of NSCs depends on the activation of PI3K/Akt pathway,but PI3K/Akt pathway may have little function in the differentiation of NSCs.
Key concepts: Wortmannin, PI3K/AKT/mTOR pathway, Protein kinase B, Phosphatidylinositol, LY294002, TUNEL assay, Blot, Apoptosis