Detection of ROS Generated by UV-C Irradiation of CdS Quantum Dots and their Effect on Damage to Chromosomal and Plasmid DNA
Detection of ROS Generated by UV-C Irradiation of CdS Quantum Dots and their Effect on Damage to Chromosomal and Plasmid DNA
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DOI：
10.1002/elan.201700648
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发表时间：
2018-04-01
期刊：
影响因子：
3
通讯作者：
Labuda, Jan
中科院分区：
文献类型：
--
作者：
Blaskovicova, Jana;Sochr, Jozef;Labuda, Jan
不同类型的DNA对损伤和突变的不同敏感性需要研究化学和物理因素对个体DNA结构完整性的影响。包括量子点在内的光活化纳米粒子引起的氧化应激被认为是其潜在风险的主要机制。以活性氧（ROS）、4-氯-7-硝基苯并-2-氧杂-1，3-二唑（NBD-Cl）和二苯基苦肼（DPPH）为指示剂，电化学DNA基生物传感器与硫醇封端的CdS量子点（CdS量子点）固定在染色体（ct DNA）和质粒（pUC 19）DNA生物识别层和琼脂糖凝胶电泳使用（i）检测CdS量子点在UV-C照射（= 254 nm）下产生的ROS，以及（ii）评估和评价它们对染色体和质粒dsDNA结构的影响。伏安法和阻抗法测定表明，在60 ~ 2400 s的照射时间内，玻碳电极表面的DNA发生了深度降解，pUC 19和ct DNA的存活率逐步下降，分别为47%和32%，与没有CdS量子点时的58%和40%相比，琼脂糖凝胶电泳支持对质粒和染色体DNA的损伤，证明双链断裂。已发现DNA类型对ROS降解的稳定性降低，顺序为pUC 19>小牛胸腺>鲑鱼精子dsDNA。
Different sensitivity of various types of DNA to damage and mutation requires a study of effects of chemical and physical factors on integrity of individual DNA structures. Oxidation stress induced by photoactivated nanoparticles including quantum dots is considered to be the main mechanism of their potential risks. In this work, spectrophotometric indicators of reactive oxygen species (ROS) 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) and -diphenyl--picrylhydrazyl (DPPH), electrochemical DNA-based biosensors with the thiol-capped CdS quantum dots (CdS QDs) immobilized over the chromosomal (ct DNA) and plasmid (pUC19) DNA biorecognition layers and agarose gel electrophoresis were used (i) to detect ROS generated at the UV-C irradiation (=254nm) of CdS QDs and (ii) to assess and evaluate their effect towards the chromosomal and plasmid dsDNA structure. Voltammetric and impedimetric measurements revealed a deep degradation of DNA attached to the glassy carbon electrode surface with a stepwise decrease of the portion of survived pUC19 and ct DNA after 60s to 2400s irradiation time up to 47% and to 32%, resp., comparing to the portion in the absence of CdS QDs reaching 58% and 40%, resp. Agarose gel electrophoresis supported the damage to plasmid and chromosomal DNA demonstrating double strand breaks. A decrease in the stability of DNA types towards degradation by ROS in order pUC19>calf thymus>salmon sperm dsDNA has been found.