Agonists of proteinase-activated receptor 2 induce TNF-α secretion from astrocytoma cells

Proteinase-activated receptor 2 (PAR2) is cleaved and activated by trypsin or mast cell tryptase, and may play an important role in inflammation. We have investigated the potential of PAR2 agonists to modulate TNF-α secretion from human astrocytoma cell line CCF-STTG1. We found that CCF-STTG1 expresses PAR2 by RT-PCR and Western blot analysis. Agonists such as trypsin, the peptide SLIGKV-NH₂ (corresponding to the PAR2 tethered ligand), or mast cell tryptase directly signal to CCF-STTG1 to stimulate secretion of TNF-α but do not stimulate in the presence of soybean trypsin inhibitor (SBTI) or VKGILS-NH₂ (reverse peptide). The secretion of TNF-α by trypsin was significantly blocked by pretreatment with either 50 μM PD98059 or 1 μM SB203580. Furthermore, trypsin stimulated the activation of extracellular signal-regulated kinase (ERK) and p38 MAP kinase homologue in CCF-STTG1 without any detectable activation of c-Jun N-terminal kinase (JNK). These results show that trypsin may induce TNF-α secretion following activation of ERK and p38 via PAR2 in CCF-STTG1.