Objective To investigate the effects of trigonelline （Trig） on the regulation of the sphingosine kinase 1 （SPHK1）/sphingosine-1-phosphate （S1P）/sphingosine-1-phosphate receptor 3 （S1PR3） pathway and its impact on the proliferation and tumorigenicity of endometrial cancer （EC） cells. Methods Ishikawa cells were divided into the following groups： The blank group， groups treated with different concentrations of Trig （low-Trig and high-Trig groups）， the SPHK1 group， the high-Trig + empty vector group， and the high-Trig + SPHK1 group. Cell proliferation was detected by colony formation assay and CCK-8 assay. The expression level of SPHK1 mRNA was measured by qRT-PCR. Western blot analysis was performed to detect the expression of antigen ki67， Myc proto-oncogene （c-Myc）proteins and proteins related to the SPHK1/S1P/S1PR3 pathway. A nude mouse xenograft tumor model was established. The tumors were excised and weighed. Western blotting was used to measure the expressions of ki67， c-Myc， and proteins related to the SPHK1/S1P/S1PR3 pathway in the xenograft tumor tissues. Hematoxylin and eosin （HE） staining and immunohistochemistry were used to detect the pathological changes of the xenograft tumor tissues and the expressions of ki67 and c-Myc， respectively. Results Compared with the blank group， the clone number， optical density （OD）， expression levels of ki67， c-Myc， SPHK1 mRNA and protein， as well as the levels of S1P and S1PR3 were lower in both the Trig-low group and the Trig-high group， while these parameters were higher in the SPHK1 group （P < 0.05）. Compared with the Trig-high + empty group， the clone number， OD， expression levels of ki67， c-Myc， SPHK1 mRNA and protein， and the levels of S1P and S1PR3 were higher in the Trig-high + SPHK1 group （P < 0.05）. Compared with the SPHK1 group， the clone number， OD， expression levels of ki67， c-Myc， SPHK1 mRNA and protein， and the levels of S1P and S1PR3 were lower in the Trig-high + SPHK1 group （P < 0.05）. In vivo experiments demonstrated that different doses of Trig could reduce the increase in the weight of transplanted tumors and inhibit the expression of proliferation-related proteins and proteins related to the SPHK1/S1P/S1PR3 pathway （P < 0.05）. Conclusions Trig effectively inhibits the proliferation of EC cells and suppresses tumor growth. The underlying mechanism is closely associated with the inhibition of the SPHK1/S1P/S1PR3 pathway.