title: Investigation of carvedilol-evoked Ca²+ movement and death in human oral cancer cells.
abstract: The effect of carvedilol on cytosolic free Ca²⁺ concentrations ([Ca²⁺](i)) in OC2 human oral cancer cells is unknown. This study examined if carvedilol altered basal [Ca²⁺](i) levels in suspended OC2 cells by using fura-2 as a Ca²⁺-sensitive fluorescent probe. Carvedilol at concentrations between 10 and 40 µM increased [Ca²⁺](i) in a concentration-dependent fashion. The Ca²⁺ signal was decreased by 50% by removing extracellular Ca²⁺. Carvedilol-induced Ca²⁺ entry was not affected by the store-operated Ca²⁺ channel blockers nifedipine, econazole, and SK&F96365, but was enhanced by activation or inhibition of protein kinase C. In Ca²⁺-free medium, incubation with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin did not change carvedilol-induced [Ca²⁺](i) rise; conversely, incubation with carvedilol did not reduce thapsigargin-induced Ca²⁺ release. Pretreatment with the mitochondrial uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) inhibited carvedilol-induced [Ca²⁺](i) release. Inhibition of phospholipase C with U73122 did not alter carvedilol-induced [Ca²⁺](i) rise. Carvedilol at 5-50 µM induced cell death in a concentration-dependent manner. The death was not reversed when cytosolic Ca²⁺ was chelated with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA/AM). Annexin V/propidium iodide staining assay suggests that apoptosis played a role in the death. Collectively, in OC2 cells, carvedilol induced [Ca²⁺](i) rise by causing phospholipase C-independent Ca²⁺ release from mitochondria and non-endoplasmic reticulum stores, and Ca²⁺ influx via protein kinase C-regulated channels. Carvedilol (up to 50 μM) induced cell death in a Ca²⁺-independent manner that involved apoptosis.
authors: Department of Dentistry, Kaohsiung Veterans General Hospital, Taiwan.
authors: Yao-Dung
authors: YD
authors: Hsieh
authors: Chao-Chuan
authors: CC
authors: Chi
authors: Chiang-Ting
authors: CT
authors: Chou
authors: Jin-Shiung
authors: JS
authors: Cheng
authors: Chun-Chi
authors: CC
authors: Kuo
authors: Wei-Zhe
authors: WZ
authors: Liang
authors: Ko-Long
authors: KL
authors: Lin
authors: Li-Ling
authors: LL
authors: Tseng
authors: Chung-Ren
authors: CR
authors: Jan