NS-398 (NS398)

NS-398 (NS398) inhibits COX-2 enzyme activity in a concentration dependent manner, the IC50 being 3.8 μM, whereas this compound at 100μM has no effect on COX-1 activity^[1]. At 10 μM, it treatment results in increased production of COX-2 and the pro-inflammatory cytokine. This compound (10 μM) induces apoptosis in LNCaP cells, but not in the more aggressive, androgen-unresponsive C4-2b cells. The C4-2b cells are observed to continue to proliferate when treated with NS-398 and continues to retain malignant phenotype characteristics. Its treatment results in C4-2b cell differentiation into an unusual neuroendocrinelike cell. These neuroendocrine-like cells produces both epithelial (cytokeratin 18 and prostate specific antigen) and neuronal (neuron-specific enolase and chromogranin A) proteins. Furthermore, this C4-2b cellular response to it is mediated by NF-kB transcription factor activation. It induces NF-kB and down-regulates Ikβ-α protein expression in LNCaP C4-2b cells^[2].

1×10_⁶ cells are plated in six well cluster plates with 2 ml of medium for 24 h. At time point 0, medium was removed, cells were carefully washed with phosphate buffered saline (PBS) and serum free(SF), phenol-red free medium containing 0.5 μg/ml BSA was added. Incubations are continued for an additional 72 h with or without increasing concentrations of NS-398 (NS398). Cells and culture medium are harvested at 24 h time intervals. Dead cells are removed by gentle washing with PBS and cell number determined by direct counting using trypan blue dye exclusion to identify viable cells. DMSO (0.1%) is added to control cultures.

NS398 (NS-398) could inhibit Cox-2 expression induced by acoustic injury and attenuate noise-induced hearing threshold shifts and cochlear hair cell loss. The inhibition of Cox-2 by this compound could attenuate Noise-induced hearing loss（NIHL）and related hair cell damage.^[3].