To detect the expression levels and clinical significance of lymphoid enhancer-binding factor 1 (LEF1) and β-catenin in eutopic and ectopic endometrium tissues of women with adenomyosis. Methods: 89 women with adenomyosis after hysterectomy were selected as the research objects from January 2019 to March 2020. The tissues of eutopic endometrium of the women were included in group A and the tissues of ectopic endometrium were included in group B, respectively. During the same time, the tissues of normal endometrium from 86 patients with cervical carcinoma were selected in group C. Real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression levels of mRNA of LEF1 and β-catenin, and immunohistochemistry was used to detect the expression levels of protein of LEF1 and β-catenin protein. Pearson method was used to analyze the correlation between LEF1 mRNA level and β-catenin mRNA level in the endometrium tissues of the women with adenomyosis, and the relationship between the levels of LEF1 mRNA and β-catenin mRNA in the endometrium tissues of these women, and their clinical features. Results: The expression level of mRNA and positive rates of protein of LEF1 and β-catenin in group B were the highest, followed by those in group A, and which in group C were lowest (P＜0.05). The expression levels of LEF1 mRNA and β-catenin mRNA in the eutopic and ectopic endometrium tissue of women with adenomyosis were positively correlated (r=0.475, 0.658, P＜0.05). There were significant difference in the uterine size and adenomyosis status between the women with positive expression of LEF1 and β-catenin in the eutopic and ectopic endometrium tissue and the women with negative expression of LEF1 and β-catenin in the eutopic and ectopic endometrium tissue (P<0.05). Conclusion: The expression levels of LEF1 and β-catenin in the ectopic and eutopic endometrium of the women with adenomyosis increase, and their abnormal expression may be related to the pathogenesis of adenomyosis.