Pten基因对小鼠胚胎成纤维细胞中Cu／Zn-SOD基因表达的调控  

Pten gene regulates the expression of Cu/Zn superoxide dismutase gene in mouse embryonic fibroblasts

作　　者：苟巧[1,2] 米粲[1] 胡迎春[2] 李刚[2] 吴德昌[2] 霍艳英[2] 

机构地区：[1]重庆医科大学基础医学院病理学教研室,重庆400016 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《第三军医大学学报》2009年第12期1127-1130,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30770652);北京市自然科学基金(7052056)~~

摘　　要：目的研究Pten基因对抗氧化蛋白Cu/Zn-SOD基因表达的调控,初步探索其调控机制。方法运用North-ern blot比较Pten+/+ MEFs与Pten-/- MEFs细胞中基础Cu/Zn-SOD mRNA表达差异,以及0.1mmol/LH2O2诱导后Pten+/+ MEFs与Pten-/- MEFs细胞中Cu/Zn-SOD mRNA表达差异;运用Western blot分析PI3K/AKT通路抑制剂LY294002作用Pten-/- MEFs细胞不同时间(30min,1、2、6、24h)后,与空白Pten-/- MEFs细胞相比,磷酸化AKT及Cu/Zn-SOD蛋白表达变化;利用Northern blot分析LY294002作用Pten-/- MEFs细胞30min,1、2、6、24h后,与空白Pten-/- MEFs细胞相比,Cu/Zn-SOD mRNA表达变化。结果Pten-/- MEFs细胞中,基础Cu/Zn-SOD mRNA表达水平降低,H2O2诱导的Cu/Zn-SOD mRNA表达明显受到抑制;在LY294002作用Pten-/- MEFs细胞30min时,AKT磷酸化水平明显降低,2h时基本没有表达;与此对应,在LY294002作用Pten-/- MEFs细胞30min时,Cu/Zn-SOD蛋白及mRNA表达均增强,并持续增强至24h。结论小鼠胚胎成纤维细胞中,Pten基因可通过拮抗PI3K/AKT信号通路调控Cu/Zn-SOD蛋白及mRNA的表达。Objective To study the regulatory effect of phosphatase and tensin homolog deleted on chro- mosome ten （Pten） gene on the expression of Cu/Zn superoxide dismutase （Cu/Zn-SOD） gene and to explore the possible mechanisms in mouse embryonic fibroblasts. Methods Northern blot was employed to measure the basal level of Cu/Zn-SOD mRNA and the expression of Cu/Zn-SOD mRNA after treatment with 0.1 mmol/L H2O2 in Pten＋/＋ and Pten-/- MEFs. Western blot was used to analyze the protein levels of phospho-AKT and Cu/Zn-SOD in Pten -/- MEFs treated with 30 txmol/L LY294002 for 0, 0.5, 1, 2, 6, and 24 h. Northern blot was applied to examine the mRNA level of Cu/Zn-SOD in Pten -/- MEFs treated with 30 μmol/L LY294002 for 0, 0.5, 1,2, 6, and 24 h. Results In Pten -/- MEFs, the basal level of Cu/Zn-SOD mRNA decreased; the 0. 1 mmol/L HeO2-indueed expression of Cu/Zn-SOD mRNA was inhibited, and the protein level of phospho- AKT decreased at 0.5, 1, 2, 6, and 24 h after treatment with 30 μmol/L LY294002. Correspondingly, the protein and mRNA levels of Cu/Zn-SOD increased at 0.5, 1, 2, 6, and 24 h after treatment with 30 μmol/L LY294002. Conclusion Pten gene may regulate the expression of Cu/Zn-SOD gene through the inhibition of the PI3 K/AKT signal pathway in mouse embryonic fibroblasts.

关 键 词：PTEN 超氧化物歧化酶 基因表达 AKT 信号通路 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学] R394-33[医药卫生—基础医学]