title: Effect of clotrimazole on cytosolic Ca(2+) rise and viability in HA59T human hepatoma cells.
abstract: Abstract Clotrimazole is an antimycotic imidazole derivative that interferes with cellular Ca(2+) homeostasis. This study examined the effect of clotrimazole on cytosolic Ca(2+) concentrations ([Ca(2+)](i)) and viability in HA59T human hepatoma cells. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)](i). Clotrimazole induced [Ca(2+)](i) rises in a concentration-dependent manner. The response was reduced by removing extracellular Ca(2+). Clotrimazole-evoked Ca(2+) entry was suppressed by store-operated channel inhibitors (nifedipine, econazole and SK&F96365) and protein kinase C modulators (GF109203X and phorbol, 12-myristate, 13-acetate). In Ca(2+)-free medium, incubation with the endoplasmic reticulum Ca(2+) pump inhibitor 2,5-di-tert-butylhydroquinone abolished clotrimazole-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 abolished clotrimazole-induced [Ca(2+)](i) rise. At 10-40 µM, clotrimazole inhibited cell viability, which was not reversed by chelating cytosolic Ca(2+). Clotrimazole at 10 and 30 µM also induced apoptosis. Collectively, in HA59T cells, clotrimazole-induced [Ca(2+)](i) rises by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via store-operated Ca(2+) channels. Clotrimazole also caused apoptosis.
authors: Department of Ophthalmology, Kaohsiung Armed Force General Hospital, Kaohsiung, Taiwan.
authors: Chi-Ting
authors: CT
authors: Horng
authors: Ni-Na
authors: NN
authors: Chiang
authors: I-Li
authors: IL
authors: Chen
authors: Wei-Zhe
authors: WZ
authors: Liang
authors: I-Shu
authors: IS
authors: Chen
authors: Daih-Huang
authors: DH
authors: Kuo
authors: Po-Chuen
authors: PC
authors: Shieh
authors: Chung-Ren
authors: CR
authors: Jan
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003022:Clotrimazole / Q000494:pharmacology*; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D016257:Fura-2; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D009543:Nifedipine / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D011493:Protein Kinase C; D010738:Type C Phospholipases; D003022:Clotrimazole; D009543:Nifedipine; D002118:Calcium; D016257:Fura-2