Effect of Notch-1 gene uprugulation on AKT-m TOR signal of U251 cell
- 医学院－已发表论文 
目的:观察上调Notch-1基因对人胶质瘤U251细胞生学物行为及AKT-m TOR信号通路的影响。方法:采用p NL-NICD慢病毒感染U251细胞,以RT-PCR和Western-Blot检测Notch-1基因mRNA和蛋白的表达,以MTT检测细胞增殖能力,流式细胞术检测细胞周期变化,Transwell实验检测细胞侵袭能力,同时检测Notch-1上调对AKT、m TOR、P70S6K、4Ebp-1蛋白的影响。结果:与对照组比较,NICD慢病毒转染72h后,Notch-1基因mRNA和蛋白表达明显增高(P<0.01);Notch-1上调明显促进U251细胞增殖、侵袭、促进细胞进入S期,增加周期蛋白Cyclin D1、CDK-4的表达,促进AKT、m TOR蛋白磷酸化。结论:上调Notch-1基因促进U251细胞增殖、侵袭,其机制和活化AKT-m TOR通路有关。Objective: To investigate the effect of Notch- 1 gene up- regulation on biological hehaviors of U215 cell and explore its effects on AKT- m TOR signal. Methods: U251 cells were infected with over- expression NICD or control lentivirus,the mRNA and protein expression of Notch- 1 gene were examined with RT- PCR and Western-Blot,the cell proliferation was assayed with MTT,cell cycle was determined by flow cytometry,the effects of Notch- 1gene on U251 cell invasion were studied using matrigel coated Transwell chambers,the effects of Notch- 1 up- gulation on AKT,m TOR,P70S6 K and 4Ebp- 1 protein expression were also analyzed. Results: Compared with the untreated and empty group,after 72 h of infection,the protein and mRNA expression of Notch- 1 increased( P < 0. 05).It was found that up- regulation of Notch- 1 promoted U251 cell growth,invasion and S- phase cells were increased( P < 0. 05). Up- regulation of Notch- 1 promoted Cyclin D1,CDK- 4 protein expression,and p- AKT and p-m TOR also increased in Notch- 1 up- regulated U251 cells. Conclusion: Up- regulation Notch- 1 promotes U251 glioma cell proliferation and invasion,the mechanism was partly through regulating AKT- m TOR signaling.