Objective: To investigate the effects of mitogen-activated protein kinase (MAPK) signaling pathway on proliferation, invasion and migration of human endometrial cancer cell line HEC-1-B. Methods: Human endometrial cancer cell line HEC-1-B was cultured in vitro. The proliferation of HEC-1-B cells was detected by MTT assay. The cell cycle of HEC-1-B was detected by flow cytometry. Transwell assay detects the invasive ability of HEC-1-B cells. The scratch test examined the migration ability of HEC-1-B cells. Western blot analysis of p-p38MAPK, PCNA and MMP-2 protein expression levels in cells. Results: p38MAPK signaling pathway inhibitor SB203580 inhibited the proliferation of endometrial cancer cell line HEC-1-B, and the inhibitory effect increased with the increase of concentration (P<0.05). Compared with the control group, the application of the inhibitor SB203580 resulted in an increase in the proportion of G0/G1 phase HEC-1-B cells, a decrease in the proportion of cells in the S phase and the G2/M phase, an increase in the inhibition rate of cell invasion, and a decrease in the rate of cell scratch healing. The expression of p-p38MAPK, PCNA and MMP-2 protein was down-regulated, and the difference was statistically significant (P<0.05). The difference between the inhibitors of SB203580 was statistically significant (P<0.05). Conclusion: Inhibition of MAPK signaling pathway can inhibit the proliferation, invasion and migration of human endometrial carcinoma HEC-1-B cells, and its mechanism may be related to down-regulation of PCNA and MMP-2 protein expression.