PTEN缺失对小鼠成纤维细胞Cu／Zn超氧化物歧化酶表达的影响及其意义  

Pilot study of PTEN deletion affecting the expression of Cu/Zn SOD

作　　者：杨柳[1] 苟巧[1] 米粲[1] 

机构地区：[1]重庆医科大学病理学教研室,400016

出　　处：《中华病理学杂志》2008年第7期477-480,共4页Chinese Journal of Pathology

摘　　要：目的探讨PTEN缺失是否影响小鼠成纤维细胞Cu／Zn超氧化物歧化酶（SOD）的表达水平及其意义。方法采用Westernblot验证及检测对照永生化小鼠胚胎成纤维细胞系PTEN^＋/＋及PTEN缺失细胞系PTEN^-/-内PTEN、磷酸化Akt、Cu／ZnSOD表达水平；Northernblot检测细胞内Cu／ZnSOD表达水平；单细胞碱性电泳检测细胞DNA损伤程度；荧光探针标记检测细胞内超氧阴离子水平；MTT法检测H2O2对细胞增殖状况的影响。结果PTEN缺失细胞系PTEN^-/-中，Cu／ZnSOD表达下调，Akt激酶磷酸化水平上调，细胞内超氧阴离子水平明显增高，空白组及H2O2处理后DNA损伤程度均较PTEN^＋/＋细胞重，H2O2对其细胞增殖的抑制作用减弱。结论PTEN缺失细胞内Cu／ZnSOD表达下调，并由此引起了细胞内活性氧持续高水平和氧化损伤累积以及活性氧对细胞增殖的抑制作用减弱。Objective To investigate whether the deletion of PTEN affects the expression of Cu/Zn SOD and the related biology. Methods Protein and mRNA expression levels of PTEN, P-Akt, Cu/Zn SOD in the control immortalized wild type mouse embryonic fibroblast cells （ PTEN^＋/＋ ） and PTEN-null cells （ PTEN^-/- ） were evaluated by Western blot and Northern blot respectively. The level of superoxide anions were detected using fluorescent probes. The DNA damage was documented by single cell alkalescence gel assay. MTT was used to study the effect of H2O2 on the proliferation of cells. Results The expression of Cu/Zn SOD was down regulated at both protein and mRNA levels, and the level of superoxide anions increased in the PTEN-null cells （ PTEN^-/- ）. The phosphorylation level of Akt kinase was up-regulated and the antiproliferative effect of H2O2 decreased in PTEN^-/- cells. Furthermore, DNA damage was observed significantly severer in both the blank control and H2O2 treated groups than that in the PTEN^＋/＋ cells. Conclusions Deletion of PTEN affects the expression of Cu/Zn SOD. As a result, reactive oxygen species （ROS） keep at a high level, along with decrease of accumulated oxidative damage and the antiproliferative effect of ROS.

关 键 词：PTEN磷酸水解酶 超氧化物歧化酶 细胞 培养的 

分 类 号：R686[医药卫生—骨科学]