title: Luciferase reporter cells as a platform to detect SMAD-dependent collagen production.
abstract: Aberrant collagen production can lead to many diseases such as fibrosis. Current methods of collagen detection are insensitive, time-consuming and laborious. We have developed a rapid, sensitive assay using chemiluminescence-based reporter cell system. Stable 3T3/NIH-SMAD-luciferase cells were generated for detection of collagen expression through TGF-β signaling, a major fibrogenic pathway. We demonstrated that these reporter cells could be used as a rapid screening tool for detection of SMAD-dependent collagen production with higher sensitivity than existing assays. Flexibility of this cell-based assay in different detection platforms makes it attractive for high throughput screening of potential fibrogenic agents and drug candidates.
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, Morgantown, WV 26506, USA.
authors: Warangkana
authors: W
authors: Lohcharoenkal
authors: Department of Computer Science and Electrical Engineering, West Virginia University, 395 Evansdale Drive, Morgantown, WV 26506, USA.
authors: Yuxin
authors: Y
authors: Liu
authors: Pathology and Physiology Research Branch, National Institute for Occupational Safety and Health (NIOSH), 1095 Willowdale Road, Morgantown, WV 26506, USA.
authors: Liying
authors: L
authors: Wang
authors: Department of Chemical Engineering, West Virginia University, 395 Evansdale Drive, P.O. Box 6102, Morgantown, WV 26506, USA.
authors: Yong
authors: Y
authors: Yang
authors: Department of Pharmaceutical Sciences, West Virginia University, 1 Medical Center Drive, Morgantown, WV 26506, USA. Electronic address: yrojan@hsc.wvu.edu.
authors: Yon
authors: Y
authors: Rojanasakul
mesh_terms: D000818:Animals; D003094:Collagen / Q000032:analysis* / Q000096:biosynthesis*; D005355:Fibrosis / Q000378:metabolism; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D015398:Signal Transduction; D051785:Smad Proteins / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D051785:Smad Proteins; D016212:Transforming Growth Factor beta; D003094:Collagen; D008156:Luciferases
keywords: Collagen; Fibrosis; Luciferase; Reporter cells; TGF-β