【摘要】 目的:研究PI3K信号通路对胃癌BGC-823细胞中P27与磷酸化P27（p-P27）蛋白表达分布的影响。方法:LY294002处理BGC-823细胞,流式细胞术测定处理前后两组细胞周期分布。Western blotting测定两组细胞总蛋白、胞浆蛋白、核蛋白中P27与p-P27（Thr187、Thr157及Ser10）蛋白含量及分布特点。结果:处理后G₁期细胞数明显增加（83.9%vs.67.6%,P<0.01）P27在总蛋白、胞浆蛋白和核蛋白中表达均显著增加（P<0.01,P<0.01,P<0.01）;p-P27（Ser10）在总蛋白、胞浆蛋白和核蛋白中表达均显著下降（P<0.01,P<0.01,P<0.01）;p-P27（Thr157）在总蛋白、胞浆蛋白中表达均显著下降（P<0.01,P<0.01）,在核蛋白中下降不明显（P=0.482）;p-P27（Thr187）在总蛋白、胞浆蛋白中表达均下降,在核蛋白中表达增高,均无显著性差异（P=0.254,P=0.70,P=0.223）。结论:阻断PI3K通路可增加胃癌细胞P27蛋白表达,降低p-P27蛋白表达,改变P27和p-P27蛋白的核浆分布,可以使细胞周期停滞于G₁期,诱导细胞凋亡。更多还原

【Abstract】 Objective:To investigate the effects of PI3K pathway inhibition on the expression and distribution of P27 and phosphorylated P27（p-P27 ） in the BGC-823 gastric cancer cell line.Methods:The cell line BGC-823 was cultivated and treated with LY294002,a PI3K pathway inhibitor.The cell cycle was determined using flow cytometry.The total cell,cytoplasmic,and nuclear expression of P27,p-P27（Thrl87 ）,p-P27（Thrl57 ）,and p-P27（SerlO ） were analyzed by Western blot analysis.Results:Inhibition of the PI3K pathway arrested cell cycle progression in the G₁ phase（treated cells vs.control cells was 83.9%vs.67.6%,P < 0.01 ）.The total cell,cytoplasmic,and nuclear expression of P27 was significantly increased in the treated group compared with those in the control group（P < 0.01,P < 0.01,and P < 0.01,respectively ）.The total cell,cytoplasmic,and nuclear expression of p-P27（Ser10 ） was significantly decreased in the treated group compared with that in the control group（P < 0.01,P < 0.01,and P < 0.01,respectively ）. The total cell and cytoplasmic expression of p-P27（Thr157 ） was significantly decreased in the treated group compared with that in the control group（P < 0.01 and P < 0.01,respectively ）.The nuclear expression of p-P27（Thr157 ） was decreased,but it did not reach statistical significance（P = 0.482 ）.The total cell and cytoplasmic expression of p-P27（Thr187 ） was decreased but the nuclear expression was increased.However,the differences were not statistically significant（P = 0.254,P = 0.70,and P = 0.223,respectively ）.Conclusion: Inhibiting the PI3K pathway in the BGC-823 gastric cancer cell line upregulates P27 expression and downregulates p-P27 expression. Moreover,the cytoplasmic and nuclear distribution of P27 and p-P27 were altered.Inhibiting the PI3K pathway induces G₁ arrest and triggers the apoptosis of gastric cancer cells.更多还原