Abstract
A cell-based assay system for monitoring COLIA2 [α2(I) collagen gene] promoter activity was developed to determine the influence of activated COLIA2 promoter in human dermal fibroblast cells. A pLuc-COLIA2 promoter plasmid that expresses the luciferase reporter gene in response to COLIA2 promoter activity was constructed. The pLuc-COLIA2 promoter plasmid and pCI-neo plasmid containing the NPT (neomycin phosphotransferase) gene for Geneticin resistance in host cells were co-transfected into human dermal fibroblast cells. COLIA2 promoter activities were measured by luciferase reporter gene assay using a luminescence detection method. Fibroblast cell transfectants treated with TNFα (tumour necrosis factor α), known to be an inhibitor of COLIA2 promoter expression, showed a reduction of COLIA2 promoter activity in a concentration-dependent manner, whereas TGF-β (transforming growth factor-β), known to be a stimulator of COLIA2 promoter expression, increased COLIA2 activity in a concentration-dependent manner. This assay system could be used to quantitatively measure COLIA2 promoter activity in human dermal fibroblast cells and allow the screening of anti-wrinkle agents from various synthetic chemicals and natural products.
| Original language | English |
|---|---|
| Pages (from-to) | 27-31 |
| Number of pages | 5 |
| Journal | Biotechnology and Applied Biochemistry |
| Volume | 47 |
| Issue number | 1 |
| DOIs | |
| State | Published - May 2007 |
| Externally published | Yes |
Keywords
- Cell-based assay
- Human dermal fibroblast transfectant cell
- Luciferase
- Transfection
- Transforming growth factor
- Type I collagen promoter
Fingerprint
Dive into the research topics of 'A cell-based assay system for high-throughput screening of anti-wrinkle agents in human dermal fibroblast transfectant cells'. Together they form a unique fingerprint.Cite this
- APA
- Author
- BIBTEX
- Harvard
- Standard
- RIS
- Vancouver