Detection of Cyto- and Genotoxicity of Rod-Shaped Gold Nanoparticles in Human Blood Lymphocytes Using Comet-FISH
Detection of Cyto- and Genotoxicity of Rod-Shaped Gold Nanoparticles in Human Blood Lymphocytes Using Comet-FISH
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DOI：
10.1508/cytologia.80.173
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发表时间：
2015-06-01
期刊：
影响因子：
1
通讯作者：
Ali, Moustafa
中科院分区：
文献类型：
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作者：
Abo-Zeid, Mona A. M.;Liehr, Thomas;Ali, Moustafa
金纳米粒子（GNP）将被应用于癌症治疗，可以想象，通过使用GNP到人体静脉的简单注射。这将使它们在到达主要目标癌细胞之前与血流中的红色和白色血细胞接触。然而，GNP对淋巴细胞可能的细胞毒性和/或遗传毒性作用尚不清楚，因此在此进行了研究。台盼蓝拒染法测定细胞毒性。对于遗传毒性，进行了Comet和Comet-FISH（=荧光原位杂交）试验。在后一项试验中，研究了DNA损伤的两种基因标记物，即作为肿瘤抑制基因的TP 53和作为肿瘤坏死因子基因的TNF-α。在不同浓度的直径为50 nm或30 nm的聚乙二醇包被的棒状GNP存在下孵育细胞。GNP在人淋巴细胞中诱导细胞毒性作用。这种影响可以以浓度和尺寸依赖性的方式观察到; 30纳米尺寸的GNPs比50纳米尺寸的GNPs毒性更大。使用彗星测定，证明了与未处理的淋巴细胞相比，GNP诱导高速率的DNA损伤，其表示为例如高比率的尾矩。靶基因（TP 53和TNF-alpha）优先在彗星尾巴中观察到，表明该DNA区域诱导的DNA损伤率很高。我们的研究结果表明，棒状GNP与人血淋巴细胞相互作用，降低细胞活力，并以浓度依赖的方式引起相关的DNA损伤。小尺寸GNP比大尺寸GNP更具有细胞毒性和遗传毒性。低浓度的大尺寸棒状GNP可能比小棒状GNP更安全地用于癌症光热治疗。但是，建议进一步研究，以尽量减少潜在的应用风险。
Gold nanoparticles (GNPs) shall be applied in cancer therapy, conceivably by using a simple injection of GNPs into human veins. This will bring them in contact with red and white blood cells of the blood stream before they reach their main target, the cancer cells. However, possible cyto- and/or genotoxic effects of GNPs on lymphocytes are not known in detail and are thus studied here. Cytotoxicity was determined by Trypan blue exclusion assay. For genotoxicity, Comet and Comet-FISH (=fluorescence in situ hybridization) assays were done. In the latter test two gene markers for DNA damage, TP53 as tumor suppressor gene and TNF-alpha as tumor necrosis factor gene, were investigated. The cells were incubated in the presence of different concentrations of polyethylene glycol coated rod-shaped GNPs of 50 nm or 30 nm in diameter. GNPs induced cytotoxic effects in human lymphocytes. The effects could be observed in concentration- and size-dependent manner; 30 nm sized GNPs were more toxic than 50 nm sized ones. Using the comet assay, it was demonstrated that GNPs induce high rates of DNA damage, which are represented e.g. as high ratios of tail moments, compared to non-treated lymphocytes. The target genes (TP53 and TNF-alpha) were observed preferentially in comet tails indicating high rates of induced DNA damage in this DNA area. Our results suggest that rod-shaped GNPs interact with human blood lymphocytes, reduce cell viability and cause relevant DNA damage in a concentration dependent manner The small sized GNPs were more cyto- and genotoxic than big sized GNPs. The low concentration of big sized rod-shaped GNPs could be safe for cancer photothermal therapy rather than small rod GNPs. However, further investigations are recommended to be able to minimize potential risks of application.