Lysophosphatidylcholine (LPC), a bioactive lipid, regulates a wide array of biological processes. LPC could be deacylated to form glycerophosphocholine by neuropathy target esterase (NTE)/Swiss cheese protein (SWS). Although NTE/SWS is important in maintaining Ca 2+ homeostasis, the role of LPC in regulating the intracellular calcium concentration ([Ca 2+ ] i ) in Drosophila remains poorly understood. We aimed to study the mechanism of LPC-induced [Ca 2+ ] i changes in Drosophila S2 cells.The [Ca 2+ ] i of Drosophila S2 cells was measured by fluorescence spectrophotometry after loading the cells with the calcium-sensitive fluorescent probe Fura-2/AM.Our results demonstrated that LPC could cause a rapid, dose-dependent increase in the [Ca 2+ ] i in the presence of external calcium ([Ca 2+ ] e ). The LPC-induced [Ca 2+ ] i increase was reduced by 60.7% in the absence of [Ca 2+ ] e . Furthermore, the Ca 2+ influx was inhibited by 37.3% after the cells were preincubated with an L-type Ca 2+ channel blocker. In the Ca 2+ -free medium, the LPC-induced [Ca 2+ ] i increase was completely blocked using an inositol triphosphate receptor (IP 3 R) inhibitor. However, a ryanodine receptor (RyR) inhibitor had no effect on the LPC-induced [Ca 2+ ] i increase.The LPC-induced [Ca 2+ ] i increase in S2 cells was dependent on both the release of Ca 2+ stored in the endoplasmic reticulum and [Ca 2+ ] e influx. Both L-type Ca 2+ channels and IP 3 R might be involved in this process. The LPC-induced [Ca 2+ ] i increase in S2 cells characterized in this study may shed light on the study of NTE/SWS protein function in general because the enzyme is responsible for the deacylation of LPC.