Screening Method for Nonsteroidal Antiinflammatory Drugs Based on the Cyclooxygenase 2 Pathway Activated by Serum-Free Stimulation in A549 Cells

抄録

  Cyclooxygenase 2 (COX-2) pathway inhibitors were regarded as promising nonsteroidal antiinflammatory drugs (NSAIDs). We discovered that the COX-2 pathway in A549 cells, a human lung cancer cell line, was activated by serum-free stimulation, and a drug screening model for NSAIDs was established based on this principle with simple performance and sufficient reliability. The COX-2 pathway was activated by treating with serum-free medium for 12 h. The activated cells were incubated with NS398 (selective COX-2 inhibitor), SC560 (selective COX-1 inhibitor), acetyl salicylic acid (ASA) (nonselective COX inhibitor) at 37°C for 15 min. Then the cells were incubated with 10 μM of arachidonic acid (AA) for another 30 min prostaglandin E₂ and 6-keto-prostaglandin F_1α were assayed in an enzyme immunoassay (EIA). The results showed that the COX-2 pathway was dominant in A549 cells whether activated by serum-free medium or not, and the COX-1 pathway could be ignored. The model accepted the positive inhibition threshold as NS398 2 μM; if a compound (10 μM) inhibited COX-2 pathway more than NS398 (2 μM), it was regarded as a hit. The COX-2 pathway inhibition experiment showed that the Z`-factor of the screening model was 0.62, which suggests that the model is suitable for COX-2 pathway inhibitor screening.