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直接纳米材料-DNA 接触对 DNA 的影响及诱变作用。

Direct nanomaterial-DNA contact effects on DNA and mutation induction.

机构信息

Department of Physics and Materials Science, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand; Thailand Center of Excellence in Physics, Commission on Higher Education, 328 Si Ayutthaya Road, Bangkok 10400, Thailand; Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand.

Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand; Biotechnology Unit, University of Phayao, Muang, Phayao 56000, Thailand.

出版信息

Toxicol Lett. 2014 Apr 7;226(1):90-7. doi: 10.1016/j.toxlet.2014.01.036. Epub 2014 Feb 3.

DOI:10.1016/j.toxlet.2014.01.036
PMID:24503012
Abstract

The toxicity of nanomaterials has been well known, but mechanisms involved have been little known. This study was aimed at looking at direct interaction between nanomaterials and naked DNA for some fundamental understanding. Two different types of nanomaterials, carbon nanotubes (CNTs) and tungsten trioxide (WO₃) nanoplates, were simply mixed with naked DNA plasmid, respectively, in two different contact modes, dry or wet (in solution), for varied time periods. DNA topological forms were analyzed for changes using gel electrophoresis and fluoro-spectrometry. The nanomaterial-contacted DNA was transferred into bacteria Escherichia coli (E. coli) cells for mutation observation. Certain types and degrees of DNA damage were observed, such as single strand break and double strand break, and bacterial mutation was confirmed. The DNA damage increased with the contacting time in an exponential manner and increased more rapidly in the initial stage for the wet contact. The nanomaterials-contacted DNA transferred bacteria had about less than 10% survival but almost 100% mutation for the surviving cells. The CNTs were more offensive than the metal oxide nanomaterials. The mutation spectrum from the DNA sequencing analysis showed that DNA point mutation was dominated by transversion, which was dominated by guanine changes in the wet contact condition while by cytosine changes in the dry contact condition. The point mutation occurrence in the wet contact was more than in the dry contact, confirming the wet contact more active and thus dangerous than dry contact. This experiment, although as a model study, revealed that direct simple contacts between nanomaterials and DNA could cause DNA changes and thus induce mutations which might potentially lead to cancers, diseases and genetic changes. This could be a mechanism for nanomaterial genotoxicity to the cells and also provided a caution to applications in using nanomaterials for DNA delivery.

摘要

纳米材料的毒性已经广为人知,但涉及的机制却知之甚少。本研究旨在研究纳米材料与裸露 DNA 之间的直接相互作用,以获得一些基本的认识。两种不同类型的纳米材料,碳纳米管(CNTs)和三氧化钨(WO₃)纳米板,分别以干接触(干燥)或湿接触(溶液)两种不同的接触方式,简单地与裸露的 DNA 质粒混合,接触时间长短不一。使用凝胶电泳和荧光光谱法分析 DNA 拓扑结构的变化。将纳米材料接触的 DNA 转移到细菌大肠杆菌(E. coli)细胞中,观察突变。观察到了某些类型和程度的 DNA 损伤,如单链断裂和双链断裂,并证实了细菌突变。DNA 损伤随接触时间呈指数增加,湿接触初期增加更快。接触纳米材料的 DNA 转移细菌的存活率约为 10%,但存活细胞的突变率几乎为 100%。CNTs 比金属氧化物纳米材料更具攻击性。DNA 测序分析表明,从突变谱来看,DNA 点突变以颠换为主,湿接触条件下以鸟嘌呤变化为主,干接触条件下以胞嘧啶变化为主。湿接触中的点突变发生多于干接触,证实湿接触比干接触更活跃,因此更危险。虽然这项实验只是一项模型研究,但它揭示了纳米材料与 DNA 之间的直接简单接触可能导致 DNA 变化,并由此引发突变,从而可能导致癌症、疾病和遗传变化。这可能是纳米材料对细胞遗传毒性的一种机制,也为纳米材料在 DNA 传递中的应用提供了警示。

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