中国药理学与毒理学杂志Issue(2)：272-276,5.DOI:10.3867/j.issn.1000-3002.2015.02.014
过氧化氢对NIH/3T3细胞蛋白磷酸酶2A催化亚基C甲基化的影响
Effect of hydrogen peroxide on protein phosphatase 2A catalytic subunit C methylation in NIH/3T3 cells
摘要
Abstract
OBJECTIVE To investigate the effect of hydrogen peroxide(H2 O2 ) on protein phospha-tase 2A catalytic subunit C(PP2Ac) methylation. METHODS After H2O2 0, 0.1, 1, 5, 10 and 25 mmol.L-1 treatment of NlH/ 3T3 cells for 1 h, the rate of resazurin reduction was observed to determine the cell proliferation while Western blotting was used to detect the expression of un-methylated PP2Ac. PP2A methylesterase (PME-1) inhibitor AMZ30 0, 0.1, 0.5, 1, 5 and 10 μmol.L-1 and antioxidant NAC 0, 0.5, 1 and 5 mmol.L-1 were used to intervene in the effect of H2 O2 1 mmol.L-1 on NlH/ 3T3 cells. After various treatments, the expression of un-methylated PP2Ac was detected by Western blotting and indirect immunofluorescence. RESULTS Compared with normal control group, H2 O2 0.1, 1, 5 and 25 mmol.L-1 significantly decreased the cell proliferation ( P < 0.01). The expression of un-methylated PP2Ac in NlH/ 3T3 cells and in H2 O2 0.1, 1 and 5 mmol.L-1 groups was significantly increased(P<0.01) compared with normal control group. Western blotting results revealed that both NAC (1 or 5 mmol.L-1 ) and AMZ30 (5 or 10 μmol.L-1 ) significantly antagonized the effect of H2 O2 on PP2Ac(P<0.01). lndirect immunofluorescence results showed that the un-methylated PP2Ac expression of H2 O2 1 mmol.L-1 group was higher than that of normal control group, while the expression of PP2Ac methylation was reduced after the use of AMZ30 10 μmol.L-1 or NAC 1 mmol.L-1. CONCLUSION H2 O2 can induce demethylation of PP2Ac in NlH/ 3T3 cells, while the PME-1 inhibitor AMZ30 and antioxidant NAC can antagonize the methylation of PP2Ac induced by H2 O2 .关键词
过氧化氢/磷蛋白磷酸酶/甲基化Key words
hydrogen peroxide/phosphoprotein phosphatase/methylation分类
医药卫生引用本文复制引用
刘银品,唐深,陆彩玲,秦富,孙斌,许扬,李习艺..过氧化氢对NIH/3T3细胞蛋白磷酸酶2A催化亚基C甲基化的影响[J].中国药理学与毒理学杂志,2015,(2):272-276,5.基金项目
国家自然科学基金(81360428)广西壮族自治区自然科学基金(2014GXNSFAA118188)@@@@The project supported by National Natural Science Foundation of China(81360428) and Natural Science Foundation of Guangxi Zhuangzu Autonomous Region(2014GXNSFAA118188) （81360428）
