The aim of this study was to detect the differential expression of Artemin (ARTN) and matrix metallopeptidase protein 9 (MMP-9) in endometrial carcinoma (EC) and to assess their clinical significance in order to provide insight into the pathological mechanism of tumor infiltration and metastasis in EC. A total of 48 patients who had undergone surgery for EC at the School of Medicine and Affiliated Hospital of HeBei University of Engineering between July 2015 and July 2016 were included in the study. The 48 patients were classified into 3 groups according to tumor stage: 27 patients with EC stage I, 12 patients with EC stage II and 9 patients with EC stage III. The samples collected from each patient included fresh normal endometrial tissue, endometrial simple hyperplastic tissue and endometrial atypical hyperplastic tissue. The transcription levels of ARTN and MMP-9 mRNA in each group were investigated using RT-PCR. The expression levels of ARTN and MMP-9 protein in each group were examined using Western blotting. Spearman's correlation analysis was used to analyze the correlation between the expression levels of ARTN and MMP-9 proteins and EC tissue type. RT-PCR and Western blotting assays revealed that the expression levels of ARTN and MMP-9 were increased in normal endometrial tissue, simple hyperplastic tissue, atypical hyperplastic tissue and EC of stages I, II and III. The differences noted were statistically significant (P<0.05). Furthermore, Western blot analysis indicated that the expression levels of ARTN and MMP-9 proteins in lymphatic metastatic tissues were higher than those in non-lymphatic metastatic tissues (P<0.05). The expression levels in the infiltration tissues of the deep muscular layer were higher than those noted in the light muscular layer (P<0.05). The expression levels of ARTN and MMP-9 proteins were positively correlated (P<0.05). The data suggest that ARTN and MMP-9 are involved in the occurrence, development, invasion and metastasis of EC, and play a synergistic role in the development of EC and lymphatic metastasis.