Evans M G, Trosko J E
Department of Pathology, Michigan State University, East Lansing.
Cell Biol Toxicol. 1988 Jun;4(2):163-71. doi: 10.1007/BF00119243.
Inhibition of gap junction-mediated cell-cell communication might be a mechanism for several types of cellular dysfunctions, including tumor promotion. Although many different assays have been designed to measure gap junction-mediated intercellular communication, we applied a new technique, termed Fluorescence Redistribution After Photobleaching ("FRAP"), to assess the ability of a known tumor promoter, 2,2', 4,4', 5,5'-hexabromobiphenyl (245-HBB), to inhibit cell-cell communication in a concentration-dependent manner. WB-F344 (rat epithelial) cells were plated at low density, exposed to noncytotoxic concentrations of 1, 5, or 20 micrograms 245-HBB/ml medium, and stained with 6-carboxyfluorescein diacetate. Single cells in pairs or clusters of touching cells in each exposure group were examined with FRAP. The results revealed an inverse correlation between the degree of fluorescence redistribution in photobleached cells and the concentration of 245-HBB. Therefore, FRAP appears to be a sensitive and rapid technique for determining complete or partial inhibition of chemically induced intercellular communication in vitro. These results also provide further evidence for the ability of 245-HBB to inhibit gap junction-mediated cell-cell communication in a concentration-dependent manner.
间隙连接介导的细胞间通讯的抑制可能是包括肿瘤促进在内的几种细胞功能障碍的一种机制。尽管已经设计了许多不同的检测方法来测量间隙连接介导的细胞间通讯，但我们应用了一种新技术，称为光漂白后荧光再分布（“FRAP”），来评估一种已知的肿瘤促进剂2,2',4,4',5,5'-六溴联苯（245-HBB）以浓度依赖性方式抑制细胞间通讯的能力。将WB-F344（大鼠上皮）细胞低密度接种，暴露于1、5或20微克245-HBB/毫升培养基的无细胞毒性浓度下，并用6-羧基荧光素二乙酸酯染色。用FRAP检查每个暴露组中配对的单个细胞或接触细胞簇。结果显示，光漂白细胞中荧光再分布的程度与245-HBB的浓度呈负相关。因此，FRAP似乎是一种灵敏且快速的技术，用于确定体外化学诱导的细胞间通讯的完全或部分抑制。这些结果也为245-HBB以浓度依赖性方式抑制间隙连接介导的细胞间通讯的能力提供了进一步的证据。
