Effect of sertraline on [Ca2+]i and viability of human MG63 osteosarcoma cells

The antidepressant, sertraline, has been shown to have diverse in vitro effects.This study examined whether sertraline altered [Ca2+], in MG63 human osteosarcoma cells by using fura-2 as a Ca2+-sensitive fluorescent dye. At 50-200 mu M, sertraline induced a [Ca2+](i) rise in a concentration-dependent manner. Ca2+ response was decreased by removing extracellular Ca2+, suggesting that Ca2+ entry and release contributed to the [Ca2+](i) signal. Sertraline-induced Ca2+ entry was inhibited by nifedipine, La3+, Gd3+, and SK&F96365. When extracellular Ca2+ was removed, pretreatment with the endoplasmic reticulum (ER) Ca2+ pump inhibitor, thapsigargin, or 2,5-di-tert-butylhydroquinone (BHQ) abolished the sertraline-evoked [Ca2+](i) rise. Incubation with sertraline also abolished the thapsigargin or BHQ-induced [Ca2+](i) rise. Inhibition of phospholipase C (PLC) with U73122 abolished the sertraline-induced [Ca2+](i) rise. At 20-30 mu M, overnight treatment with sertraline killed cells in a concentration-dependent manner. The cytotoxic effect of sertraline was not reversed by chelating cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA).Annexin V/propidiunn iodide staining data demonstrate that sertraline (30 mu M) evoked apoptosis. Sertraline (20 and 30 mu M) also increased levels of reactive oxygen species.Together, in human osteosarconna cells, sertraline evoked a [Ca2+](i) rise by inducing PLC-dependent Ca2+ release from the ER and Ca2+ entry by L-type Ca2+ channels and store-operated Ca2+ channels. Sertraline induced cell death that may involve apoptosis by mitochondrial pathways.