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| DNA氧化损伤修复基因hOGG1高表达细胞株的建立 | |
| 引用本文： | 王昱,卢仲毅,许峰,朱宇熹,许峰.DNA氧化损伤修复基因hOGG1高表达细胞株的建立[J].第二军医大学学报,2010,31(7):725-730. |
| 作者姓名： | 王昱 卢仲毅 许峰 朱宇熹 许峰 |
| 作者单位： | 1. 第三军医大学西南医院儿科,重庆,400038;重庆医科大学附属儿童医院PICU,重庆,400014 2. 重庆医科大学附属儿童医院PICU,重庆,400014 3. 重庆医科大学附属第一医院肿瘤科,重庆,400016;Department of Radiation Oncology & Image-applied Therapy, Kyoto University Graduate School of Medicine, 606-8225 Japan |
| 基金项目： | 国家自然科学基金面上项目，重庆市自然科学基金 |
| 摘 要： | 目的联合pcDNA3.1(+)/Myc-HisA-hOGG1和pGL3promoter荧光质粒稳定转染人肺腺癌A549细胞获得hOGG1基因高表达细胞株,并初步探讨转染细胞生物学特性的变化。方法成功构建pcDNA3.1(+)/Myc-His A载体后,大量抽提阳性重组子并在Fu GENE6的介导下联合pGL3promoter荧光质粒转染A549细胞(转染组),同时设空白对照组(A549)和阴性对照组PGL3+pcDNA3.1(+)/Myc-HisA转染的A549细胞]。生物荧光检测转染细胞hOGG1mRNA表达的改变,蛋白质免疫印迹法检测转染细胞hOGG1蛋白表达水平。将3组细胞于不同高氧条件下培养,相差显微镜观察形态学变化,彗星试验比较各组细胞对抗损伤和修复能力的情况,同时测定DNA氧化损伤标志物8-羟基脱氧鸟苷(8-OHdG)的变化。结果转染细胞荧光素酶稳定表达,蛋白印迹检测转染组hOGG1蛋白明显高于两对照组细胞,提示hOGG1基因高表达细胞株建立成功。相同高氧条件下,转染组形态学变化明显减轻,彗星细胞率和olive tail moment明显低于空白组和阴性对照组,修复完成率高,修复时间缩短(P<0... |
| 关 键 词： | hOGG1基因 共转染 彗星试验 DNA损伤修复 |
| 收稿时间： | 2010/1/13 0:00:00 |
| 修稿时间： | 2010/5/17 0:00:00 |
Establishment of a lung cancer cell line A549 highly expressing DNA oxidative damage related repair gene hOGG1 |
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| WANG Yu,LU Zhong-yi,XU Feng,ZHU Yu-xi,and xufeng.Establishment of a lung cancer cell line A549 highly expressing DNA oxidative damage related repair gene hOGG1[J].Academic Journal of Second Military Medical University,2010,31(7):725-730. | |
| Authors: | WANG Yu LU Zhong-yi XU Feng ZHU Yu-xi and xufeng |
| Institution: | WANG Yu1,2,LU Zhong-yi2,XU Feng2,ZHU Yu-xi3,41.Department of Pediatrics,Southwest Hospital,Third Military Medical University,Chongqing 400038,China2.Department of PICU,the Affiliated Pediatric Hospital of Chongqing Medical University,Chongqing 400014,China 3.Department of Oncology,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China 4.Department of Radiation Oncology & Image-applied Therapy,Kyoto University Graduate School of Medicine,606-8225 Japan |
| Abstract: | Objective To establish a human lung cancer cell line A549 highly expressing human 8-oxoguanine-DNA glycosylase(hOGG1)by co-transfecting pcDNA 3.1(+)/Myc-HisA-hOGG1 and PGL3 promoter,and to observe the biological behavior of the transfected cells.Methods PcDNA3.1(+)/Myc-His A-hOGG1 and PGL3 promoter were steadily co-transfected into A549 cells via mediation of Fu GENE 6(transfected group);untransfected cells served as blank control and cells transfected with PGL3+pcDNA3.1(+)/Myc-HisA served as negative contr... |
| Keywords: | hOGG1 co-transfection comet assay DNA damage and repair |
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