This study aimed to investigate the expression of Six1 gene and its downstream target gene Ezrin in cervical cancer, and to correlate their expression to the clinical pathology of cervical cancer. RT-PCR and Western blot were used to detect the expression of Six1 and Ezrin in cervical cancer tissue, cervical intraepithelial neoplasia tissue, and normal cervical tissue. The correlation of Six1 and Ezrin expression with the occurrence, development, and clinical pathology of cervical cancer was then analyzed. The expression of Six1 and Ezrin mRNA and protein in cervical cancer and cervical intraepithelial neoplasia was significantly higher than in normal cervical tissue (p < 0.05). Their expression was also significantly higher in the cancerous tissues of patients with advanced cervical cancer than in those of patients with early-stage cervical cancer (p < 0.05). Moreover, they were expressed at significantly higher levels in lymph node metastasis-positive patients than in lymph node metastasis-negative patients (p < 0.05). Finally, we observed that lesser differentiated tumors had elevated expression of Six1 and Ezrin mRNA and protein (p < 0.05). However, the mRNA and protein expression of Six1 and Ezrin were independent of patient age and tumor size (p > 0.05). Further investigation is warranted to describe the relation of whether Six1 and Ezrin protein contributes to the mechanism of occurrence, development, differentiation, and invasiveness of the tumor. These markers could be used as indicators of prognosis in early-stage cervical cancer as Six1 and Ezrin had a synergistic effect on the incidence of cervical cancer.