Pten基因敲除对过氧化物酶家族表达和活性氧水平的影响

Effect of Pten gene knock-out on peroxiredoxins expression and reactive oxygen species levels

目的:探讨Pten基因敲除后对过氧化物酶家族(Peroxiredoxins,Prdxs)水平和活性氧水平的影响。方法:采用Western印迹和化学/荧光发光分析法分别检测了在Pten+/+MEF和Pten-/-MEF细胞中PRDXs的表达和细胞内活性氧水平。结果:Western印迹结果显示,与Pten+/+MEF细胞相比,Pten-/-MEF细胞PRDXⅠ,Ⅱ,Ⅴ,Ⅵ蛋白水平下调,PRDXⅢ不变,PRDXⅣ上调。DCFH探针标记后流式结果显示Pten-/-MEF细胞活性氧荧光值显著高于对照Pten+/+MEF细胞(P<0.05)。结论:Pten基因敲除引起数种PRDXs表达下调,细胞内活性氧水平增高。

Objective： To analyze the effect of Pten gene knock-out on peroxiredoxins（PRDXs） expression and the reactive oxygen species （ROS） levels in cells. Methods： The PRDXs protein level and the ROS levels of control Pten^＋/＋ MEF cells and Pten^ -/- MEF cells were assessed by Western blot and fluorescence activated cell sorther（FACS） method respectively. Results： Compared with control Pten^ ＋/＋ MEF cells,the protein level of PRDXⅠ,Ⅱ,Ⅴ,Ⅵ was down-regulated,the PRDX Ⅲ level did not change and the PRDX1V level was up-regulated in Pten^ -/- MEF cells. The ROS fluorescence cell number was singnificantly higher in Pten^-/- MEF cells than in Pten^＋/＋ MEF cells. Conclusion： Pten gene knock-out down-regulates PRDX Ⅰ,Ⅱ,Ⅴ,Ⅵ expression and increases ROS levels in cell.