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目的研究10号染色体缺失磷酸酶及张力蛋白同源体(Phosphatase and tensin homolog deleted on chromosome ten,PTEN)再表达对细胞内抗氧化蛋白Prdx1,2,5,6和Cu/Zn SOD的表达、活性氧(ROS)和DNA氧化损伤水平及细胞抗氧化能力的影响。方法采用Western blot法比较Pten-/-MEFs细胞(对照细胞)与PTEN再表达的Pten-/-MEFs细胞内抗氧化蛋白Prdx1,2,5,6和Cu/Zn SOD的表达差异;采用化学/荧光发光法分析对照细胞与PTEN再表达细胞内基础ROS水平差异;采用中性单细胞凝胶电泳比较不同浓度(0、0.01、0.05、0.10mmol/L)H2O2处理后,对照细胞与PTEN再表达细胞内DNA双链断裂(DSBs)的变化。结果与对照细胞相比,PTEN再表达细胞中Prdx1,2,5,6和Cu/Zn SOD蛋白的表达水平增高,细胞内基础ROS水平降低,DSBs减少,细胞抵抗外源性H2O2的能力增强。结论 PTEN可能通过对Prdx1,2,5,6和Cu/Zn SOD等抗氧化蛋白的表达调控,增强细胞抗氧化能力,清除细胞内过量的ROS,从而保护细胞免受氧化压力导致的DNA氧化损伤,维持基因组稳定性。
Objective To study the expression and activity of anti-oxidative proteins Prdx1, 2, 5, 6 and Cu / Zn SOD in re-expression of Phosphatase and tensin homolog deleted on chromosome ten (PTEN) Oxygen (ROS) and DNA oxidative damage and cellular antioxidant capacity. Methods The expressions of Prdx1, 2, 5, 6 and Cu / Zn SOD in Pten - / - MEFs and Pten - / - MEFs were compared by Western blot. Fluorescence method was used to analyze the difference of basal ROS levels between control cells and PTEN re-expressing cells. After treated with different concentration of H2O2 (0, 0.01, 0.05, 0.10mmol / L) by neutral single cell gel electrophoresis, the control cells and PTEN were reexpressed Changes in intracellular DNA double-strand breaks (DSBs). Results Compared with the control cells, the expression of Prdx1, 2, 5, 6 and Cu / Zn SOD protein in PTEN overexpression cells increased, the intracellular ROS level decreased, the DSBs decreased and the cell resistance to exogenous H2O2 increased. Conclusion PTEN may protect cells against oxidative DNA damage by regulating the expression of anti-oxidative proteins Prdx1, 2, 5, 6 and Cu / Zn SOD, enhancing cellular anti-oxidative capacity and clearing excessive intracellular ROS. , To maintain genomic stability.