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In LY\u2010B cells, deficiency of sphingolipids enhanced the release of AA induced by bee venom sPLA<jats:sub>2<\/jats:sub>\u2010III or human sPLA<jats:sub>2<\/jats:sub>\u2010V. These alterations were reversed by replenishment of exogenous sphingomyelin (SM). In LY\u2010A cells, deficiency of SM increased the release of AA induced by sPLA<jats:sub>2<\/jats:sub>. In CHO\u2010K1 cells, decrease and increase of SM level in the plasma membrane by pharmacological methods increased and inhibited the release of AA, respectively. SM inhibited the activity of sPLA<jats:sub>2<\/jats:sub> in vitro. Niemann\u2010Pick disease type C (NPC) is a lysosomal storage disorder caused by mutation of either the NPC1 or NPC2 gene, and is characterized by accumulation of cholesterol and sphingolipids including SM in late endosomes\/lysosomes. Increased levels of AA and sPLA<jats:sub>2<\/jats:sub> activity are involved in various neurodegenerative diseases. In CHO cells lacking NPC1 (A101 cells), SM level was lower in the plasma membrane, while it was higher in late endosomes\/lysosomes. The release of AA induced by sPLA<jats:sub>2<\/jats:sub> was increased in A101 cells than that in parental cells (JP17 cells), which was attenuated by adding exogenous SM. In addition, sPLA<jats:sub>2<\/jats:sub>\u2010III\u2010induced cytotoxicity in A101 cells was much higher than that in JP17 cells. These results suggest that SM in the plasma membrane plays important roles in regulating sPLA<jats:sub>2<\/jats:sub> activity and the enzyme\u2010induced cytotoxicity in A101 cells. J. Cell. 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