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Background & Aims Secondary bile acids like deoxycholic acid (DCA) are well-established tumor promoters that may exert their pathologic actions by interfering with intracellular signaling cascades. Methods We evaluated the effects of DCA on Ca²⁺ signaling in BHK-21 fibroblasts using fura-2 and mag-fura-2 to measure cytoplasmic and intraluminal internal stores [Ca²⁺], respectively. Furthermore, green fluorescent protein (GFP)-based probes were used to monitor time courses of phospholipase C (PLC) activation (pleckstrin-homology [PH]-PLCδ-GFP), and translocation of protein kinase C (PKC) and a major PKC substrate, myristolated alanine-rich C-kinase substrate (MARCKS). Results DCA (50–250 μmol/L) caused profound Ca²⁺ release from intracellular stores of intact or permeabilized cells. Correspondingly, DCA increased cytoplasmic Ca²⁺ to levels that were ∼120% of those stimulated by Ca² …