Emerging evidence suggests that microbiota dysbiosis plays a critical role in the pathogenesis of endometrial cancer (EC), a leading cause of cancer-related deaths in women globally. However, few studies have simultaneously examined both the upper and lower genital tract microbiota in such individuals. In this study, we investigated alterations in microbiota composition across different parts of the female genital tract in a Chinese cohort of EC patients. Samples from 59 individuals (22 endometrial cancer patients; 8 endometrial hyperplasia patients, and 29 benign controls) were collected. In addition, a total of 58 vaginal swabs, 39 fallopian swabs, 16 peritoneal fluid samples, 36 urine swabs, and 34 endometrium samples were finally recruited. The composition of bacterial communities was determined by 16S ribosomal RNA Miseq sequencing. Specific taxa were significantly enriched in the EC group, including