Luciferase reporter cells as a platform to detect SMAD-dependent collagen production

J Biosci Bioeng. 2014 Dec;118(6):732-5. doi: 10.1016/j.jbiosc.2014.05.021. Epub 2014 Jun 25.

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.

Keywords: Collagen; Fibrosis; Luciferase; Reporter cells; TGF-β.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Collagen / analysis*
  • Collagen / biosynthesis*
  • Fibrosis / metabolism
  • Genes, Reporter*
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Mice
  • NIH 3T3 Cells
  • Signal Transduction
  • Smad Proteins / metabolism*
  • Transforming Growth Factor beta / metabolism

Substances

  • Smad Proteins
  • Transforming Growth Factor beta
  • Collagen
  • Luciferases