摘要
目的探讨PI3K信号通路调控肺癌细胞增殖机制。方法培养肺癌细胞系H446和A549,LY294002特异性阻断PI3K信号通路后,MTT比色法测定细胞生长曲线;Western blot法检测PI3K信号下游蛋白SKP2、p27表达变化;FCM分析细胞周期变化。结果与对照组相比,处理组细胞生长速率受到抑制;Western blot结果显示H446、A549细胞中的SKP2蛋白表达水平降低,而p27蛋白表达水平升高;FCM结果表明抑制剂处理后肿瘤细胞阻滞于G1期。结论结果提示肺癌细胞中PI3K信号通路参与SKP2蛋白的表达调控,从而影响p27蛋白的降解,促进细胞周期,加速细胞增殖。
Objective To investigate the mechanism of PI3K signaling-mediated proliferation of lung cancer cells. Methods The human lung cancer cell lines H446 and A549 were cultured and treated with specific PI3K inhibitor LY294002 at 25 μmol/L to block the PI3K pathway. MTr colorimetric assay, flow cytometer （FCM） and western blot were employed to plot the cell growth curves, analyze the cell cycles and measure the expression of SKP2 and p27 proteins which were the downstream effectors of PI3K pathway. Results Compared with control group, the cellular growth of the LY294002-treated was significantly inhibited. Western blotting revealed that inhibition of PI3K signal- ing decreased the expression of SKP2 protein,while increased the expression of p27 protein. FCM showed that PI3K inhibitor could arrest the cell cycle to G1 phase. Conclusion These data implicate that PI3K signaling might regulate the cell cycle and proliferation of lung cancer cells by modulating the SKP2-dependent degradation of p27.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2009年第9期663-665,共3页
Journal of China Medical University
基金
辽宁省教育厅科学研究计划项目(2004D225)
