Mitochondrial localization of superoxide dismutase is required for decreasing radiation-induced cellular damage
Mitochondrial localization of superoxide dismutase is required for decreasing radiation-induced cellular damage
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DOI：
10.1667/rr3081
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发表时间：
2003-11-01
影响因子：
3.4
通讯作者：
Greenberger, JS
中科院分区：
文献类型：
--
作者：
Epperly, MW;Gretton, JE;Greenberger, JS
我们调查的重要性线粒体定位的SOD 2（MnSOD）转基因产品的保护32 D Cl 3造血细胞辐射诱导的杀伤。四个质粒含有（1）天然人铜/锌超氧化物歧化酶（Cu/ZnSOD，SOD 1）转基因，（2）天然SOD 2转基因，（3）减去线粒体定位前导序列的SOD 2转基因（MnSOD-ML），和（4）与SOD 1转基因的活性部分连接的SOD 2线粒体前导序列（ML-Cu/ZnSOD）转染32 Dcl 3细胞，经卡那霉素抗性筛选获得亚克隆株。每个细胞克隆的克隆形成体外辐射存活曲线显示，与亲本系32 D cl 3（D-o 1.15 +/- 0.11戈伊）相比，表达Cu/ZnSOD和MnSOD-ML的克隆的细胞辐射抗性没有增加（D-o分别= 0.89 +/- 0.01和1.08 +/- 0.02戈伊）。相反，表达SOD 2或ML-Cu/ZnSOD的细胞克隆具有显著的辐射抗性（D-o分别= 2.1 +/- 0.1和1.97 +/- 0.17戈伊）。与对照照射小鼠或腹腔注射Cu/ZnSOD-PL或MnSOD-ML-PL的小鼠相比，腹腔注射SOD 2-质粒-脂质体（MnSOD-PL）复合物的小鼠在35戈伊后表现出显著较少的食管炎。在一项实验中，食管内注射ML-Cu/ZnSOD-PL的小鼠显示出显著的辐射防护作用。这些数据表明，在体外和体内保护细胞免受辐射诱导的细胞损伤的SOD的线粒体定位的重要性。(C)2003年，辐射研究协会。
We investigated the importance of mitochondrial localization of the SOD2 (MnSOD) transgene product for protection of 32D cl 3 hematopoietic cells from radiation-induced killing. Four plasmids containing (1) the native human copper/zinc superoxide dismutase (Cu/ZnSOD, SOD1) transgene, (2) the native SOD2 transgene, (3), the SOD2 transgene minus the mitochondrial localization leader sequence (MnSOD-ML), and (4) the SOD2 mitochondrial leader sequence attached to the active portion of the SOD1 transgene (ML-Cu/ZnSOD) were transfected into 32D cl 3 cells and subclonal lines selected by kanamycin resistance. Clonogenic in vitro radiation survival curves derived for each cell clone showed that Cu/ZnSOD- and MnSOD-ML- expressing clones had no increase in cellular radiation resistance (D-o = 0.89 +/- 0.01 and 1.08 +/- 0.02 Gy, respectively) compared to parent line 32D cl 3 (D-o 1.15 +/- 0.11 Gy). In contrast, cell clones expressing either SOD2 or ML-Cu/ZnSOD were significantly radioresistant (D-o = 2.1 +/- 0.1 and 1.97 +/- 0.17 Gy, respectively). Mice injected intraesophageally with SOD2-plasmidAiposome (MnSOD-PL) complex demonstrated significantly less esophagitis after 35 Gy compared to control irradiated mice or mice injected intraesophageally with Cu/ZnSOD-PL or MnSOD-ML-PL. Mice injected with intraesophageal ML-Cu/ZnSOD-PL showed significant radioprotection in one experiment. The data demonstrate the importance of mitochondrial localization of SOD in the in vitro and in vivo protection of cells from radiation-induced cellular damage. (C) 2003 by Radiation Research Society.