RRC ID |
43464
|
著者 |
Hojo H, Igawa K, Ohba S, Yano F, Nakajima K, Komiyama Y, Ikeda T, Lichtler AC, Woo JT, Yonezawa T, Takato T, Chung UI.
|
タイトル |
Development of high-throughput screening system for osteogenic drugs using a cell-based sensor.
|
ジャーナル |
Biochem Biophys Res Commun
|
Abstract |
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.
|
巻・号 |
376(2)
|
ページ |
375-9
|
公開日 |
2008-11-14
|
DOI |
10.1016/j.bbrc.2008.08.167
|
PII |
S0006-291X(08)01725-7
|
PMID |
18789892
|
MeSH |
Alkaline Phosphatase / analysis
Alkaline Phosphatase / biosynthesis
Animals
Biosensing Techniques / methods*
Bone Morphogenetic Protein 2
Bone Morphogenetic Proteins / pharmacology
Cell Line
Collagen Type I / genetics
Drug Evaluation, Preclinical / methods*
Fluorescence
Genes, Reporter
Green Fluorescent Proteins / analysis
Green Fluorescent Proteins / genetics
Isoflavones / pharmacology
Mice
Osteoblasts / drug effects*
Osteocalcin / analysis
Osteocalcin / biosynthesis
Osteogenesis / drug effects*
Osteoporosis / drug therapy*
Rats
Transforming Growth Factor beta / pharmacology
|
IF |
2.985
|
引用数 |
14
|
WOS 分野
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
ヒト・動物細胞 |
MC3T3-E1(RCB1126) |