Akt抑制剂MK2206对膀胱癌细胞增殖凋亡的影响

Effect of Akt Inhibitor-MK2206 on Proliferation and Apoptosis of T24 Cells

目的探讨Akt抑制剂MK2206对膀胱癌细胞(T24)增殖、凋亡的影响及其作用机制。方法采用CCK-8法检测不同时间、不同浓度(0,1,5,10,20μmol·L-1) MK2206对T24细胞增殖的影响,流式细胞术检测不同浓度MK2206对T24细胞凋亡的影响; Western blotting检测Akt磷酸化蛋白表达情况及GSK-3β/β-catenin通路蛋白的表达水平。结果 CCK-8法检测显示,MK2206对T24细胞增殖活力具有显著的抑制效果(P<0.05),且在0～20μmol·L-1内呈浓度-时间依赖性。流式细胞术检测结果表明,MK2206在作用T24细胞24 h后,能够促进T24细胞凋亡。Western blotting检测显示,p-Akt(ser347)蛋白表达量显著降低(P<0.01);同时,MK2206在对GSK-3β总蛋白表达无明显的影响,P-GSK-3β、β-catenin蛋白表达水平显著降低(P<0.05)。结论 MK2206能够抑制T24细胞增殖,促进其发生凋亡,从而缓解膀胱癌发展进程,其作用机制可能是抑制GSK-3β/β-catenin信号通路。

Objective To explore the effect and mechanism of Akt inhibitor MK2206 on proliferation and apoptosis of T24 cells. Methods The CCK-8 method was used to detect the cell proliferation. Then the effects of different concentrations (0,1,5,10,20 μmol·L ^-1 ) MK2206 on T24 cell apoptosis were tested by flow cytometry. Mealwhile, Western blotting was used to detect the phosphorylation of Akt protein and GSK-3β/β-catenin pathway proteins. Results The result of CCK-8 assay showed that MK2206 had a significant inhibitory effect on the proliferation activity of T24 cells ( P <0.05), and the effect was in time-dose dependence manner arranged 0-20 μmol· L^-1 concentration. Flow cytometry results showed that the MK2206 could promote apoptosis of T24 cells in 24 h. Comparing to control group, the level of p-Akt (ser347) protein significantly decreased ( P < 0.01). There was a significantly inhibitory effect on the GSK-3β/β-catenin signaling pathway, which showed the expression of p-GSK-3β and β-catenin proteins was significantly reduced ( P <0.05) in the premise of no influence on the expression of GSK-3β. Conclusion MK2206 has inhibited T24 cell proliferation and promoted cell apoptosis via restraining Akt phosphorylation (ser473) and inhibiting the GSK-3β/β-catenin signaling pathway. MK2206 may relieve the tumor development process.