Effect of thymol on Ca super(2+ homeostasis and viability in human glioblastoma cells)

The effect of the natural essential oil thymol on cytosolic Ca super(2+ concentrations ([Ca) super(2)+] sub(i) and viability in human glioblastoma cells was examined. The Ca) super(2)+-sensitive fluorescent dye fura-2 was applied to measure [Ca super(2+]) sub(i). Thymol at concentrations of 400-1000 mu M induced a [Ca super(2+]) sub(i) rise in a concentration-dependent fashion. The response was decreased partially by removal of extracellular Ca super(2+. Thymol-induced Ca) super(2)+ signal was not altered by nifedipine, econazole, SK&F96365, and protein kinase C activator phorbol myristate acetate (PMA), but was inhibited by the protein kinase C inhibitor GF109203X. When extracellular Ca super(2+ was removed, incubation with the endoplasmic reticulum Ca) super(2)+ pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) abolished thymol-induced [Ca super(2+]) sub(i) rise. Incubation with thymol also abolished thapsigargin or BHQ-induced [Ca super(2+]) sub(i) rise. Inhibition of phospholipase C with U73122 abolished thymol-induced [Ca super(2+]) sub(i) rise. At concentrations of 200-800 mu M, thymol killed cells in a concentration-dependent manner. This cytotoxic effect was not changed by chelating cytosolic Ca super(2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl (BAPTA/AM). Annexin V/propidium iodide staining data suggest that thymol (200, 400 and 600 mu M) induced apoptosis in a concentration-dependent manner. Collectively, in human glioblastoma cells, thymol induced a [Ca) super(2)+] sub(i rise by inducing phospholipase C- and protein kinase C-dependent Ca) super(2)+ release from the endoplasmic reticulum and Ca super(2+ entry via non store-operated Ca) super(2)+ channels. Thymol induced cell death that may involve apoptosis.