inositol-1-4-5-trisphosphate and Hepatoblastoma

Previously, we have reported that inhibition of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channels by glibenclamide induced intracellular Ca2+ release from IP(3)-sensitive stores and apoptosis in HepG2 human hepatoblastoma cells (Kim JA, Kang YS, Lee SH, Lee EH, Yoo BH, Lee YS. 1999. Biochem Biophys Res Commun 261:682-688). In this study we investigated the upstream signals involved in the mechanism of these actions of glibenclamide. Treatment with glibenclamide initiated production of inositol 1,4,5-trisphosphate (IP(3)) in a dose- and time-dependent manner. The glibenclamide-induced formation of IP(3) was significantly inhibited by CFTR activators (levamisole and bromotetramisole). The intracellular Ca2+ release and apoptosis induced by glibenclamide were significantly suppressed by treatment with phospholipase C (PLC) inhibitors (U-73122 and manoalide) or by pretreatment with pertussis toxin (PTx). In addition, PTx-catalyzed ADP-ribosylation of GTP-binding proteins (G-proteins) was markedly enhanced by treatment with glibenclamide in a time-dependent manner. Taken together, these results suggest that PTx-sensitive G-proteins coupled to PLCbeta may mediate the intracellular Ca2+ release and apoptosis induced by inhibiting CFTR Cl- channels in HepG2 cells. These results further suggest that the PTx-sensitive G-proteins may be a valuable target for the therapeutic intervention of human hepatomas.

Topics: Adenosine Diphosphate Ribose; Apoptosis; Calcium; Catalysis; Cystic Fibrosis Transmembrane Conductance Regulator; Glyburide; GTP-Binding Proteins; Hepatoblastoma; Inositol 1,4,5-Trisphosphate; Liver Neoplasms; Pertussis Toxin; Signal Transduction; Tumor Cells, Cultured; Type C Phospholipases; Virulence Factors, Bordetella