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基于细胞阻抗谱技术的微囊藻毒素-LR 和节球藻毒素对细胞抑制作用的无创探测。

Noninvasive probing of inhibitory effects of cylindrospermopsin and microcystin-LR using cell-based impedance spectroscopy.

机构信息

Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, Canada H4P 2R2.

出版信息

Environ Sci Technol. 2010 Sep 1;44(17):6775-81. doi: 10.1021/es101206t.

Abstract

In an effort to develop a noninvasive method for assessment of cyanobacterial toxins in drinking water, plausible cytotoxicity/inhibition of microcystin-LR and cylindrospermopsin was evaluated by cell-substrate impedance sensing (ECIS) using three different cell lines. Sf9 insect cells were attached to concanavalin A coated gold electrodes, whereas Chinese hamster ovary (CHO) and human embryo kidney (HEK) cells were attached to a fibronectin or laminin coated gold surface. Cytotoxic or inhibitory effects were dependent upon the cell line and the extracellular matrix (ECM) coating. Neither toxin exhibited any appreciable effect on the insect cells. In contrast, cytotoxicity of cylindrospermopsin on CHO cells was attested by both ECIS and viability tests. The half-inhibition concentration (ECIS50) of cylindrospermopsin for CHO cells was approximately 2 microg/mL (ppm) after 20 h of exposure and 4 microg/mL (ppm) after 30 h of exposure for a laminin or fibronectin coated surface. ECIS confirmed no significant effect of cylindrospermopsin on HEK cells. Microcystin-LR was also tested with CHO cells, resulting in an ECIS50 value of approximately 12 microg/mL (ppm) after 25 h of exposure for a laminin coated gold surface. The effect of microcystin-LR on CHO cells probed by ECIS was inhibitory rather than cytotoxic, as confirmed by cell viability assays.

摘要

为了开发一种非侵入性的方法来评估饮用水中的蓝藻毒素,使用细胞底物阻抗感应(ECIS)技术,通过三种不同的细胞系评估了微囊藻毒素-LR 和节旋螺毒素的潜在细胞毒性/抑制作用。 Sf9 昆虫细胞附着在刀豆球蛋白 A 涂覆的金电极上,而中国仓鼠卵巢(CHO)和人胚肾(HEK)细胞附着在纤维连接蛋白或层粘连蛋白涂覆的金表面上。细胞毒性或抑制作用取决于细胞系和细胞外基质(ECM)涂层。两种毒素都没有对昆虫细胞产生明显的影响。相比之下,节旋螺毒素对 CHO 细胞的细胞毒性通过 ECIS 和活力测试得到证实。暴露 20 小时后,节旋螺毒素对 CHO 细胞的半抑制浓度(ECIS50)约为 2 微克/毫升(ppm),暴露 30 小时后,用纤维连接蛋白或层粘连蛋白涂覆的表面为 4 微克/毫升(ppm)。ECIS 证实节旋螺毒素对 HEK 细胞没有显著影响。还用 CHO 细胞测试了微囊藻毒素-LR,结果显示在层粘连蛋白涂覆的金表面暴露 25 小时后,ECIS50 值约为 12 微克/毫升(ppm)。ECIS 探测到微囊藻毒素-LR 对 CHO 细胞的影响是抑制性的,而不是细胞毒性的,这通过细胞活力测定得到了证实。

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