Anchored Cell Analysis/Sorting Coupled with the Scrape-Loading/Dye-Transfer Technique to Quantify Inhibition of Gap-Junctional Intercellular Communication in WB-F344 Cells by 2,2',4,4', 5,5'-Hexabromobiphenyl

摘要

Inhibition of intercellular communication has been hypothesized to play a role in tumor promotion. The compound 2,2', 4,4', 5,5' -hexabromobiphenyl (245-HBB) is a tumor promoter in vivo and blocks inter-cellular communication in vitro. The scrape-loading/dye-transfer (SL/DT) assay was used to assess this in vitro effect at varying concentrations of 245-HBB. The SL/DT technique is based on the inter-cellular loading of a fluorescent dye, lucifer yellow (LY), and monitoring its transfer into adjacent cells via patent gap junctions. Confluent WB-F344 (rat epithelial) cells were exposed to various non-cytolethal concentrations of 245-HBB. Transfer of LY was then quantified with anchored cell analysis/sorting. The results indicate an inverse correlation between the degree of fluorescence in secondary LY-recipient cells and the treatment concentration. The coupling of these two new methods of cellular biology provided rapid quantitative analysis of dye transfer in measuring the concentration/response of modulation of gap-junctional permeability in cultured cells. Keywords: Toxicity, Reprints. (AW)