一种基于绿色荧光蛋白的蛋白酶体抑制剂细胞筛选模型

Establishment of a GFP-based cellular model for screening novel proteasome inhibitors

摘要:

为建立基于绿色荧光蛋白(GFP)的药物筛选模型, 并用此模型从包括中药提取物在内的化合物中筛选新型蛋白酶体抑制剂, 本研究构建了pGC-E1-ZU1-GFP融合蛋白慢病毒表达载体并感染A549细胞, 筛选稳定表达细胞株, 用已知蛋白酶体抑制剂PS-341处理细胞, 荧光显微镜检测处理前后细胞GFP水平变化。结果获得了稳定表达pGC-E1-ZU1-GFP的A549细胞, 这些细胞用PS-341处理24 h后用荧光显微镜检测, 发现细胞绿色荧光强度相对于对照组明显增强。利用这一模型对一些化合物进行筛查, 发现了一些新的蛋白酶体抑制剂。

Abstract:

To establish a green fluorescent protein (GFP)-based cellular model for screening proteasome inhibitors from compounds including extracts from Traditional Chinese Medicinal herbs, we transfected A549 cells with lentivirus expression vector pGC-E1-ZU1-GFP, and selected clones stably expressing ZU1-GFP. The A549-ZU1-GFP cells were treated with PS-341 for 24 h, and the accumulation of GFP was analyzed by fluorescence microscope. We found that the fluorescence intensity of A549-ZU1-GFP cells treated with PS-341 was significantly increased as compared to the control. We screened for proteasome inhibitors from compounds including some from Traditional Chinese Medicinal herbs, and the data suggested a few compounds could be novel proteasome inhibitors.