Ding Min, Bowman Linda, Castranova Vincent
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV, USA.
Methods Mol Biol. 2012;906:403-14. doi: 10.1007/978-1-61779-953-2_33.
Nanotechnology exploits the fact that nanoparticles exhibit unique physicochemical properties, which are distinct from larger particles of the same composition. It follows that nanoparticles may also express distinct bioactivity and unique interactions with biological systems. Therefore, it is essential to assess the potential health risks of exposure to nanoparticles to allow development and implementation of prevention measures. One of the biggest challenges facing the field of nanotoxicology is the huge variety of different nanoparticle types possessing a variety of properties. Genetic Luciferase Reporter System or Reporter gene assay has become an invaluable tool in studies of gene expression. This is achieved by linking the firefly luciferase gene to a promoter sequence. Luciferase assays are quick, highly sensitive, have wide dynamic range, and are cheap to perform. Because of their simplicity and versatility, and because of the absence of endogenous luciferase activity in most cell types, this test can be applied for testing a large variety of nanomaterials for their pathogenic or carcinogenetic effects on a wide range of mammalian cells. This system is an ideal early-stage toxicology tool for screening nanomaterials. Here we describe the Genetic Luciferase Reporter System as the method for detecting alteration of gene expression in response to external stimuli (e.g., nanoparticles).
纳米技术利用了纳米颗粒具有独特物理化学性质这一事实，这些性质与相同成分的较大颗粒不同。因此，纳米颗粒也可能表现出独特的生物活性以及与生物系统的独特相互作用。所以，评估接触纳米颗粒的潜在健康风险对于制定和实施预防措施至关重要。纳米毒理学领域面临的最大挑战之一是存在大量具有各种性质的不同类型纳米颗粒。基因荧光素酶报告系统或报告基因检测已成为基因表达研究中一项非常有价值的工具。这是通过将萤火虫荧光素酶基因与启动子序列连接来实现的。荧光素酶检测快速、高度灵敏、动态范围广且操作成本低。由于其简单性和通用性，以及大多数细胞类型中不存在内源性荧光素酶活性，该检测可用于测试多种纳米材料对多种哺乳动物细胞的致病或致癌作用。该系统是筛选纳米材料的理想早期毒理学工具。在此，我们将基因荧光素酶报告系统描述为检测基因表达响应外部刺激（如纳米颗粒）变化的方法。
