Effects Of MTOR Inhibitor AZD8055 On The Biological Of Cholangiocarcinoma Cells And Its Molecular Mechanism

Objectives:To explore the effects of mTOR inhibitor AZD8055 on the biological of cholangiocarcinoma cells and its molecular mechanism.Materials and methods:The effect of proliferation by treated with AZD8055 was detected by MTT assay.The colony formation of HuCCT1 cells after treated with AZD8055 was tested by colony formation assay.The effect of cell migratin by treated with AZD8055 was detected by wound healing assay.The effect of apoptosis by treated with AZD8055 was detected by flow cytometry.Western blot was used for detecting the expression of autophagy,anti-apoptosis and Akt/mTOR signaling pathway related proteins.Results:1.MTT assay results showed that compared with the control group,AZD8055 on 25nM,50nM,100nM,200nM and 400nM inhibited cell proliferation of HuCCTl following treated,and showed a dose-dependent(P<0.05),2.Colony formation assay showed that compared with the control group,after treated with 25nM,50nM and 100nM AZD8055 inhibited the colony formation following(P<0.05).3.By using wound healing assay,compared with the control group,cholangiocarcinoma cells showed significantly short transport distance after treated with 100nM and 200nM AZD8055(P<0.05).4.Flow cytometry showed that treated with AZD8055,cell apoptosis was induced weakly.5.Western blot analysis showed,compared with the control group,the expression of Beclin-1 and the ratio of LC3 II/LC3 I was enhanced.The expression of Bax and Cleaved caspase 3 was down-regulated and Bcl-2 was up-regulated.down regulationThe expression level of phosphorylated protein of Akt,S6 and 4EBP1 was significantly decreased.Conclusions:1.AZD8055 can inhibit the proliferation of cholangiocarcinoma cells,and its mechanism is related to the induction of autophagy and down-regulation of Akt/mTOR signaling pathway.2.AZD8055 can inhibit the horizontal migration of cholangiocarcinoma cells,and its mechanism is related to down-regulation of Akt/mTOR signaling pathway.