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首页> 外文会议>12th Symposium on Bioluminescence and Chemiluminescence, Apr 5-9, 2002, Robinson College, University of Cambridge, UK >APPLICATION OF A LUMINESCENCE WHOLE-CELL BIOSENSOR BASED ON A RECOMBINANT MOUSE HEPATOMA CELL LINE FOR THE DETECTION OF DIOXIN-LIKE CHEMICALS
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APPLICATION OF A LUMINESCENCE WHOLE-CELL BIOSENSOR BASED ON A RECOMBINANT MOUSE HEPATOMA CELL LINE FOR THE DETECTION OF DIOXIN-LIKE CHEMICALS

机译：基于重组小鼠肝细胞系的发光全细胞生物传感器在二恶英样化学物质检测中的应用

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摘要

Dioxin-like chemicals are a diverse group of widespread environmental contaminants, which include polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs). These polyhalogenated aromatic hydrocarbons (PHAHs) are very stable, thus persist in the environment and can be found in soil, sediment, water, air, food. They are highly toxic at low doses, accumulate in fatty tissues and milk of living organisms due to their lipophilicity and biomagnify in the food chain. Such compounds are also semi-volatile, which enables them to move long distances in the atmosphere before deposition occurs. They are known to produce a variety of species- and tissue-specific effects on humans and wildlife, such as tumor promotion, birth defects, hepatotoxicity, immunotoxicity, dermal toxicity, endocrine disruption. Current techniques for detection and quantitation of PHAHs rely on costly and time-consuming instrumental analysis methods, such as gas chromatography coupled to mass spectrometry (GC-MS), and require complex sample preparation procedures. Whole-cell biosensors represent an alternative approach which exploits the receptor/reporter concept. They are based on the use of recombinant cell lines which naturally express the aryl hydrocarbon receptor (AhR) and are genetically modified by the introduction of a plasmid bearing a reporter gene which is induced by the binding of dioxin-like chemicals to the AhR. In the present work a luminescence whole-cell biosensor based on a recombinant mouse hepatoma cell line and the firefly luciferase luc sequence as a reporter gene was characterized and optimised. The possibility of applying the biosensor to the direct analysis of milk samples was also assessed.
机译：类二恶英类化学物质是多种多样的广泛的环境污染物，包括多氯化二苯并对二恶英（PCDD），二苯并呋喃（PCDF）和联苯（PCB）。这些多卤代芳烃（PHAH）非常稳定，因此可以在环境中持久存在，并且可以在土壤，沉积物，水，空气，食物中找到。它们在低剂量下具有高毒性，由于它们的亲脂性并在食物链中被生物放大，因此会积聚在生物的脂肪组织和牛奶中。这些化合物也是半挥发性的，这使它们能够在沉积发生之前在大气中移动很长一段距离。已知它们会对人类和野生生物产生各种物种和组织特有的效应，例如肿瘤促进，先天缺陷，肝毒性，免疫毒性，皮肤毒性，内分泌破坏。当前用于检测和定量PHAH的技术依赖于昂贵且费时的仪器分析方法，例如气相色谱与质谱联用（GC-MS），并且需要复杂的样品制备程序。全细胞生物传感器代表了一种利用受体/记者概念的替代方法。它们基于重组细胞系的使用，该重组细胞系天然表达芳烃受体（AhR），并通过引入带有报告基因的质粒进行了遗传修饰，该报告基因是由二恶英样化学物质与AhR的结合诱导的。在本工作中，基于重组小鼠肝癌细胞系和萤火虫荧光素酶luc序列作为报告基因的发光全细胞生物传感器得到了表征和优化。还评估了将生物传感器应用于牛奶样品直接分析的可能性。

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