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Cell-Based Sensing of Endocrine Disrupting Substances Using Fluorescent Protein-Gold Nanoparticle ComplexesWang, Xian 29 August 2014 (has links)
Developing a sensitive and effective in vitro bioassay to detect endocrine disrupting chemicals (EDCs) would reduce the cost, eliminate the possibility of low dose effects, detect the non-monotonic dose responses, and identify mechanisms of actions. The “chemical nose” sensing method using supramolecular complexes composed of cationic monolayer functionalized gold nanoparticles (AuNPs) and fluorescent proteins (FPs) can successfully distinguish serum proteins, mammalian cells, tissue lysates, and chemotherapeutic drug mechanisms. EDCs regulate target cells via genomic or non-genomic pathways in terms of proliferative effect and response time. In this thesis, green fluorescent protein-gold nanoparticle (GFP-AuNP) sensors were used to detect the proliferative effect of 17b-estradiol (E2) and bisphenol A (BPA) on MCF7 and T47D cell lines at fM or pM dose range. Non-monotonic dose responses were also observed at different exposure times. The dose-response relationships using GFP-AuNP sensors could be correlated to the cell cycle analysis. Interestingly, tamoxifen, an estrogen antagonist, showed distinct patterns at low doses on HepG2 cells using triple channel FP-AuNP sensors, which might indicate different mechanisms of actions in this dose range.
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