Calcium is a key signaling molecule in β-lapachone-mediated cell death

beta -Lapachone (beta -Lap) triggers apoptosis in a number of human breast and prostate cancer cell lines through a unique apoptotic pathway that is dependent upon NQO1, a two-electron reductase. Downstream signaling pathway(s) that initiate apoptosis following treatment with beta -Lap have not been elucidated, Since calpain activation was suspected in beta -Lap-mediated apoptosis, we examined alterations in Ca2+ homeostasis using NQO1-expressing MCF-7 cells. beta -Lap-exposed MCF-7 cells exhibited an early increase in intracellular cytosolic Ca2+, from endoplasmic reticulum Ca2+ stores, comparable to thapsigargin exposures. 1,2-Bis-(2-aminophenoxy) ethane-N,N,N ' ,N ' -tetraacetic acid-acetoxymethyl ester, an intracellular Ca2+ chelator, blocked early increases in Ca2+ levels and inhibited beta -Lap-mediated mitochondrial membrane depolarization, intracellular ATP depletion, specific and unique substrate proteolysis, and apoptosis, The extracellular Ca2+ chelator, EGTA, inhibited later apoptotic end points (observed >8 h, e.g. substrate proteolysis and DNA fragmentation), suggesting that later execution events were triggered by Ca2+ influxes from the extracellular milieu. Collectively, these data suggest a critical, but not sole, role for Ca2+ in the NQO1-dependent cell death pathway initiated by beta -Lap. Use of beta -Lap to trigger an apparently novel, calpain like-mediated apoptotic cell death could be useful for breast and prostate cancer therapy.