Lin Chi-Iou, Hsieh Chun-Hung, Lee Stanley Shiao-Ying, Lee Wei-Shan, Chang-Chien Gou-Ping, Pan Chien-Yuan, Lee Hsinyu
Institute of Zoology, National Taiwan University, No. 1, Roosevelt Rd., Sec. 4, Taipei, 106, Taiwan, ROC.
J Biomed Sci. 2008 Nov;15(6):833-40. doi: 10.1007/s11373-008-9267-6. Epub 2008 Jul 5.
Dioxins comprise a group of compounds which contain a double aromatic ring-like structure. They are among the most prevalent and toxic environmental pollutants. Accumulation of dioxins in human tissues poses a potential threat to human health. Currently, analytical chemical procedures dominate dioxin-detection protocols. In this study, we established a fluorescence resonance energy transfer (FRET)-based dioxin-detection bioassay. Aryl hydrocarbon receptor (AHR) and AHR nuclear translocator (ARNT) fused-cyan fluorescent protein (CFP) and -yellow fluorescent protein (YFP) constructed were transiently co-transfected into rat hepatoma cell line, H4IIEC3 cells. Our results showed that no FRET signals were detected in AHR-CFP- and ARNT-YFP-transfected H4IIEC3 cells. However, dioxin treatments upregulated FRET signals in these transfected cells in a dose-dependent manner. This work highlighted the potential of FRET technique in the detection of dioxin-like compounds.
二噁英是一组含有双芳香环结构的化合物。它们是最普遍且有毒的环境污染物之一。二噁英在人体组织中的积累对人类健康构成潜在威胁。目前,分析化学程序主导着二噁英检测方案。在本研究中,我们建立了一种基于荧光共振能量转移(FRET)的二噁英检测生物测定法。将构建的芳基烃受体(AHR)和AHR核转运体(ARNT)与青色荧光蛋白(CFP)和黄色荧光蛋白(YFP)融合,瞬时共转染到大鼠肝癌细胞系H4IIEC3细胞中。我们的结果表明,在转染了AHR-CFP和ARNT-YFP的H4IIEC3细胞中未检测到FRET信号。然而,二噁英处理使这些转染细胞中的FRET信号以剂量依赖方式上调。这项工作突出了FRET技术在检测二噁英类化合物方面的潜力。