Sphingosylphosphorylcholine (SPC) has been shown to be a potent mitogen for Swiss 3T3 fibroblasts and also to be an inhibitor of cell growth of some cancer cells, suggesting cell-selective action of the lipid. We examined the effects of SPC, and its structurally-related sphingosine (SP), sphingosine 1-phosphate (S1-P) and membrane-permeable derivatives of ceramides on cell growth of four strains of human pancreatic cancer cells, MLA PaCa-2, PANC-1, PK-1 and PK-9. Under the reported conditions for SPC-induced stimulation of 3T3 fibroblasts, where cells were grown to confluency in the presence of 10% fetal bovine serum (FBS) in culture prior to experiments and insulin was supplemented in experimental culture, none of the agents tested stimulated DNA synthesis in MIA PaCa-2 cells and ceramide at high concentration even inhibited it. On the other hand, in reduced FBS concentration in preculture and in the absence of insulin in experimental culture, SP, S1-P and ceramides suppressed cell growth of all the cells tested including Swiss 3T3 fibroblasts. However, under these conditions, SPC inhibited three out of four species of pancreatic cancer cells but stimulated Swiss 3T3 fibroblasts in terms of both DNA synthesis and cell proliferation. Cell cycle analysis showed that SPC stimulated cell cycle progress from the G1 to the S phase in Swiss 3T3 fibroblasts but inhibited it in PANC-1 cells in reduced FBS concentrations. We suggest that extracellular SPC can inhibit cell growth of human pancreatic cancer cells through regulation of the cell cycle process depending upon both the cell species and environmental conditions.