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使用氧化石墨烯筛选单链DNA断裂光敏剂的快速简便方法。

Fast and simple method for screening of single-stranded DNA breaking photosensitizers using graphene oxide.

作者信息

Kim Joong Hyun, Kim Hyun Jin

机构信息

Medical Device Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu, 41061, South Korea.

出版信息

Nano Converg. 2018 Oct 22;5(1):29. doi: 10.1186/s40580-018-0160-3.

Abstract

A fast and simple method for screening of ssDNA-breaking photosensitizers was developed using graphene oxide. The ultraviolet light-induced DNA breaks are one of the most harmful DNA damages and cause skin cancer if they are left unrepaired. Since graphene oxide showed relatively strong affinity to the broken DNA than intact DNA, and it quenched fluorescence of the DNA labeling dye effectively, the degree of ultraviolet light-induced broken DNAs could be analyzed by measuring decreased fluorescence after mixing the DNA with graphene oxide. The decrease of fluorescence was highly correlated with the ultraviolet light-irradiating time and concentration of the added drugs. As a result, it was possible to evaluate the efficacy of different ssDNA-breaking photosensitizers in a high-throughput manner. However, conventional methods for the damaged-DNA analysis are time-consuming and require additional manipulations such as purification, radio-labeling, enzymatic digestion, or chemical modification of DNA. The phototoxicity of five drugs such as benzophenone, ketoprofen, indomethacin, naproxen, and norfloxacin was tested using the proposed method. The ssDNA-breaking efficiency of the drugs was well matched with reported efficiency of the tested drugs. In contrast to naked gold nanoparticles, graphene oxide is stably dispersed in the presence of salt, the phototoxicity of the drugs could be successfully tested at a physiological condition using the graphene oxide based method.

摘要

开发了一种使用氧化石墨烯筛选单链DNA断裂光敏剂的快速简便方法。紫外线诱导的DNA断裂是最有害的DNA损伤之一,如果不进行修复会导致皮肤癌。由于氧化石墨烯对断裂的DNA显示出比对完整DNA更强的亲和力,并且能有效淬灭DNA标记染料的荧光,因此通过测量DNA与氧化石墨烯混合后荧光的降低程度,可以分析紫外线诱导的断裂DNA的程度。荧光的降低与紫外线照射时间和添加药物的浓度高度相关。结果,可以高通量方式评估不同单链DNA断裂光敏剂的功效。然而,传统的受损DNA分析方法耗时且需要额外的操作,如DNA的纯化、放射性标记、酶切或化学修饰。使用所提出的方法测试了二苯甲酮、酮洛芬、吲哚美辛、萘普生和诺氟沙星等五种药物的光毒性。药物的单链DNA断裂效率与所测试药物的报道效率非常匹配。与裸金纳米颗粒不同,氧化石墨烯在盐存在下能稳定分散,使用基于氧化石墨烯的方法可以在生理条件下成功测试药物的光毒性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee94/6196149/3b0e086aaec9/40580_2018_160_Fig1_HTML.jpg

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