Identification of upregulators of BMP2 expression via high-throughput screening of a synthetic and natural compound library.
Bone morphogenetic protein II (BMP2), a member of the transforming growth factor-beta (TGF-beta) superfamily, is highly expressed in osteoblasts and is a crucial regulator of osteogenic differentiation. Many observations clearly indicate the high potency of BMP2 as an inducer of osteogenesis, and it may be a novel therapeutic target for diseases associated with bone loss, especially in menopausal and postmenopausal women. To discover new agents that enhance the expression of the mouse BMP2, the authors developed a high-throughput assay to screen a synthetic and natural compound library. The cell-based high-throughput screen was conducted in 96-well plates using the clonal murine calvarial MC3T3-E1 cells. These cells were stably transfected with mouse BMP2 promoter-luciferase and calibrated with the known antiosteoporosis compound genistein. Among 3192 compounds screened, 3 agents (daidzein, formononetin, and 2-Acetyldibenzothiophene) were picked up by the high-throughput screening assay, and those compounds were identified as upregulators of BMP2 expression by real-time quantitative reverse transcription-polymerase chain reaction and flow cytometry. Thus, it is demonstrated that this screening model is useful for identifying lead compounds to treat osteoporosis and maintain bone metabolism balance.
Li
Xue
X
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
Yang
Jun
J
He
Xiaobo
X
Yang
Zhaoyong
Z
Ding
Yan
Y
Zhao
Pei
P
Liu
Zongying
Z
Shao
Huayi
H
Li
Zhuorong
Z
Zhang
Yueqin
Y
Si
Shuyi
S
D000818:Animals; D001688:Biological Products / Q000032:analysis* / Q000494:pharmacology*; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism; D019833:Genistein / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007529:Isoflavones / Q000737:chemistry / Q000494:pharmacology; D049449:Luminescence; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects*
D001688:Biological Products; D055396:Bone Morphogenetic Protein 2; D007529:Isoflavones; D012333:RNA, Messenger; C004742:daidzein; D019833:Genistein