--- - |2- Sphingosylphosphorylcholine (SPC) could induce apoptosis through c‐Jun N‐terminal kinase (JNK) signaling in MDA‐MB‐231 cells. An antagonistic signaling about autophagy/AKT/p38 was activated to inhibit JNK signaling and subsequent apoptosis by SPC. Though evoking autophagy/AKT/p38 pathways that anatagonize apoptosis, SPC eventually led to cell apoptotic death. Abstract Sphingosylphosphorylcholine (SPC), an important lipid mediator in blood, inhibits the proliferation and migration of various cancer cells. However, its effect as a cell‐specific sphingolipid in breast cancer cells is still unknown. Here, we showed that SPC promoted autophagy and apoptosis in triple‐negative breast cancer MDA‐MB‐231 cells. Autophagy worked as a negative regulator of apoptosis‐induced by SPC. Mechanistically, SPC mediated apoptosis via activating c‐Jun N‐terminal kinase (JNK). Meanwhile, p38MAPK (p38) and protein kinase B (PKB or AKT) signaling pathways were also activated to inhibit apoptosis, suggesting that SPC could evoke multiple signaling pathways to modulate cell apoptosis. In addition, the crosstalk between autophagy, p38, AKT and JNK is that autophagy, p38, and AKT attenuated the JNK. AKT and p38 were in the downstream of autophagy, which is autophagy/AKT/p38 signaling evoked by SPC to antagonize JNK signaling and subsequent apoptosis. Although the pathways that antagonize apoptosis were evoked, the cells eventually reached apoptosis by SPC. Therefore, the combination with pharmacological autophagy inhibitors would be a more effective therapeutic strategy for eliminating breast cancer cells by SPC. - 'Journal of Cellular Physiology, EarlyView. '