Development of high-throughput screening system for osteogenic drugs using a cell-based sensor
Development of high-throughput screening system for osteogenic drugs using a cell-based sensor
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DOI：
10.1016/j.bbrc.2008.08.167
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发表时间：
2008-11-14
影响因子：
3.1
通讯作者：
Chung, Ung-il
中科院分区：
文献类型：
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作者：
Hojo, Hironori;Igawa, Kazuyo;Chung, Ung-il
为了有效治疗骨质疏松症及其他骨质流失疾病，需要能强力诱导骨形成的小分子化合物。本研究最初尝试建立一种监测系统，以便轻松、精确且无创地检测成骨分化情况。为此，我们构建了经大鼠I型胶原蛋白启动子2.3kb片段驱动的绿色荧光蛋白（GFP）报告基因稳定转染的前成骨细胞MC3T3E1（Col1a1GFP - MC3T3E1）。在这些细胞中，我们筛选出了一个在骨形态发生蛋白2（BMP2）诱导成骨刺激下会发出荧光的克隆。GFP荧光强度与碱性磷酸酶（ALP）染色强度以及骨钙素（Oc）信使核糖核酸（mRNA）水平高度相关。利用该系统，我们对天然和合成化合物库进行了筛选，从而鉴定出一种异黄酮衍生物——光甘草异黄酮（GI）。GI以剂量依赖的方式诱导ALP染色和Oc mRNA表达。Col1a1GFP - MC3T3E1系统或许有助于鉴定新型促骨生成药物。（C）2008爱思唯尔公司版权所有。
To effectively treat osteoporosis and other bone-loss disorders, small compounds that potently induce bone formation are needed. The present study initially attempted to establish a monitoring system that Could detect osteogenic differentiation easily, precisely, and noninvasively. For this purpose, we established pre-osteoblastic MC3T3E1 cells stably transfected with the GFP reporter gene driven by a 2.3 kb fragment of rat type I collagen promoter (Col1a1GFP-MC3T3E1). Among these cells, we selected a clone that fluoresced upon osteogenic stimulation by BMP2. The GFP fluorescence intensity corresponded well to the intensity of alkaline phosphatase (ALP) staining and to the level of osteocalcin (Oc) mRNA. Using this system, we screened natural and synthetic compound libraries and thus identified an isoflavone derivative, glabrisoflavone (GI). GI induced ALP staining and Oc mRNA in a dose-dependent manner. The Col1a1GFP-MC3T3E1 system may be useful for identifying novel osteogenic drugs. (C) 2008 Elsevier Inc. All rights reserved.