Abstract
Objective: To investigate the activity of cyclooxygenase-2(COX-2) gene promoter of human gastric cancer cells under inflammatory state. Methods: The expression of COX-2 in human gastric cancer tissues was detected with immunohistochemical staining. Recombinant pGL3-COX-2-promoter of human COX-2 gene promoter was constructed, and gastric cancer cell line SGC-7901 and BGC-823 were co-transfected with pGL3-COX-2-promoter and pRL-TK with lipofectin reagent-mediated transfection. Cells were collected after treatment with different mass concentrations of lipopolysaccharide (LPS) for different time, the activity of dual-luciferase reporter gene was determined with dual-luciferase reporter system, and the expression of COX-2 protein was detected by Western blotting. Results: Immunohistochemical staining indicated that COX-2 protein expressed both in cytoplasm of gastric cancer cells and cytoplasm of para-cancer mucosal epithelial cells. At the same time point, the activity of COX-2 gene promoter of SGC-7901 cells and BGC-823 cells increased with the mass concentrations of LPS(P < 0.05). The activity of COX-2 gene promoter reached the peak after treatment with high concentration of LPS(10 μg/mL) for 6 h, and then gradually decreased(P < 0.05). Western blotting revealed that the expression of COX-2 protein of SGC-7901 cells and BGC-823 cells increased with the time of treatment with 10 μg/mL LPS. Conclusion: Inflammatory stimulation may activate COX-2 gene promoter of human gastric cancer cells.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 283-287+335 |
| Journal | Journal of Shanghai Jiaotong University (Medical Science) |
| Volume | 32 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 2012 |
| Externally published | Yes |
Keywords
- Cyclooxygenase-2
- Dual-luciferase report gene
- Gastric cancer
- Lipopolysaccharide
- Promoter
ASJC Scopus subject areas
- General Biochemistry, Genetics and Molecular Biology