Autonomous multielectrode system for monitoring the interactions of isoflavonoids with lung cancer cells.
Autonomous multielectrode system for monitoring the interactions of isoflavonoids with lung cancer cells.
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DOI：
10.1021/ac035436m
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发表时间：
2004-03
影响因子：
7.4
通讯作者：
S. Andreescu;O. Sadik;D. McGee;S. Suye
中科院分区：
文献类型：
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作者：
S. Andreescu;O. Sadik;D. McGee;S. Suye
流行病学和饮食研究已经报道了异黄酮类化合物在不同形式的癌症发生中的细胞毒性作用。因此，人们正在寻找一种准确可靠的方法来监测这些化学物质与癌细胞的相互作用。为研究异黄酮类化合物对A549肺腺癌细胞株的作用，我们研制并优化了一种全自主的电化学生物传感器。这种先进的生物传感器使用了一个原型96电极(DOX-96)井型设备，允许通过消耗溶解氧来测量细胞的呼吸活动。该系统提供对暴露于自然产生的多酚，特别是白藜芦醇、染料木素和槲皮素的细胞活动的连续、实时监测。该系统配备了一个多电位仪，一个用于测量和细胞培养的96电极井，每个井装有3个一次性电极。与经典的“细胞培养”技术比较表明，该生物传感器无需添加试剂即可实现实时测量。检测下限为1×10(4)个/孔，而MTT法和荧光法分别为200和6×10(3)个/孔。该方法在细胞稳定性、重复性、应用潜力、每孔细胞密度、每孔细胞培养液的体积/组成以及培养时间等方面进行了优化。其他包括总测量时间、温度和灭菌程序。这项研究代表了一个基础研究工具，可以让研究人员研究异黄酮类化合物的类型、水平和对细胞的特定影响。
The cytotoxic effect of isoflavonoids in the development of different forms of cancer has been reported by epidemiological and dietary studies. Consequently, there is a search for an accurate and reliable method for monitoring the interactions of these chemicals with cancerous cells. We have developed and optimized a fully autonomous electrochemical biosensor for studying the role of isoflavonoids on A549 lung adenocarcinoma cell line. This advanced biosensor uses a prototype 96-electrode (DOX-96) well-type device that allows the measurement of cell respiratory activity via the consumption of dissolved oxygen. The system provides a continuous, real-time monitoring of cell activity upon exposure to naturally occurring polyphenols, specifically resveratrol, genistein, and quercetin. The system is equipped with a multipotentiostat, a 96-electrode well for measurements and cell culturing with 3 disposable electrodes fitted into each well. A comparison with classical "cell culture" techniques indicates that the biosensor provides real-time measurement with no added reagents. A detection limit of 1 x 10(4) was recorded versus 200 and 6 x 10(3) cells/well for MTT and fluorescence assays, respectively. This method was optimized with respect to cell stability, reproducibility, applied potential, cell density per well, volume/composition of cell culture medium per well, and incubation. Others include total measuring time, temperature, and sterilization procedure. This study represents a basic research tool that may allow researchers to study the type, level, and specific influence of isoflavonoids on cells.