Damaged DNA-binding protein 2 accelerates UV-damaged DNA repair in human corneal endothelium
Damaged DNA-binding protein 2 accelerates UV-damaged DNA repair in human corneal endothelium
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DOI：
10.1016/j.exer.2004.06.010
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发表时间：
2004-09-01
影响因子：
3.4
通讯作者：
Yamagami, S
中科院分区：
文献类型：
--
作者：
Inoki, T;Hitoshi, E;Yamagami, S
目的.检测体外和离体损伤的DNA结合蛋白2基因表达水平，以及紫外线照射后损伤的DNA结合蛋白2过表达的人角膜内皮细胞的DNA修复程度。通过半定量逆转录-聚合酶链反应测定了各种人体组织中组成性损伤的DNA结合蛋白2基因表达水平。采用核糖核酸酶保护试验研究了紫外线照射后培养的人角膜内皮细胞中核苷酸切除修复相关基因表达的动态变化。用携带损伤DNA结合蛋白2的腺病毒感染培养的人角膜内皮细胞，观察损伤DNA结合蛋白2对紫外线照射后角膜内皮细胞DNA修复的影响。在所研究的各种人体组织中，供体角膜中的人角膜内皮和上皮具有最高的组成性损伤DNA结合蛋白2基因表达。与核苷酸切除修复因子相关的基因表达水平动力学研究表明，紫外线照射后，培养的人角膜内皮细胞中受损的DNA结合蛋白2-基因表达的增加率最高的核苷酸切除修复相关基因的研究。体内DNA修复实验表明，过表达DNA结合蛋白2的培养人角膜内皮细胞经紫外线照射后，其DNA修复效率明显高于对照组。人角膜内皮大量表达受损的DNA结合蛋白2基因，该基因在紫外线照射下有效产生。人角膜内皮细胞中这种过表达的受损DNA结合蛋白2有助于保护系统免受紫外线照射后的DNA损伤。我们的研究结果表明，受损的DNA结合蛋白2在DNA修复中起关键作用，以维持人类角膜内皮功能。(C)2004爱思唯尔有限公司保留所有权利。
Purpose. To determine damaged DNA-binding protein 2-gene expression levels in vitro and ex vivo, and the degree of DNA repair in damaged DNA-binding protein 2-overexpressing cultured human corneal endothelium after ultraviolet irradiation.Methods. Constitutive damaged DNA-binding protein 2-gene expression levels in various human tissues were determined by semi-quantitative reverse transcription-polymerase chain reactions. The dynamics of nucleotide excision repair-related gene expression in cultured human corneal endothelium were investigated in a ribonuclease protection assay after ultraviolet-irradiation. The effect of damaged DNA-binding protein 2 on DNA repair was studied after ultraviolet-irradiation in cultured human corneal endothelium infected with adenovirus carrying damaged DNA-binding protein 2.Results. Human corneal endothelium and epithelium in the donor cornea had the highest constitutive damaged DNA-binding protein 2-gene expression of the various human tissues studied. Gene expression level dynamics associated with nucleotide excision repair factors after ultraviolet-irradiation showed that the increase in the rate of damaged DNA-binding protein 2-gene expression in cultured human corneal endothelium was highest of the nucleotide excision repair-related genes studied. An in vivo DNA repair assay showed that DNA repair efficiency in damaged DNA-binding protein 2-overexpressing cultured human corneal endothelium after ultraviolet-irradiation was significantly improved as compared with that in the control human corneal endothelium.Conclusion. The human corneal endothelium abundantly expresses the damaged DNA-binding protein 2-gene that is produced efficiently on ultraviolet exposure. This overexpressed damaged DNA-binding protein 2 in the human corneal endothelium contributes to the protection system against DNA damage after ultraviolet-irradiation. Our findings show a critical role for damaged DNA-binding protein 2 in DNA repair to maintain the human corneal endothelium function. (C) 2004 Elsevier Ltd. All rights reserved.