It has been shown that death-associated protein kinase (DAPK) family and E-cadherin play significant roles in the promotion of apoptosis and the suppression of cell adhesion and migration, and are involved in tumor metastasis. Ezrin, a cytoplasmic peripheral membrane protein, has been shown to interact with E-cadherin to participate in the metastasis of tumor cells. The present study aimed to investigate the expression of DRP-1 (a member of the DAPK family), ezrin and E-cadherin in esophageal squamous cell carcinoma (ESCC), and to analyze their association with clinicopathological factors in order to explore their potential in ESCC diagnosis. The expression of these genes was studied in tissue microarrays using in situ hybridization and immunohistochemistry methods in 76 specimens of ESCC and their paracancerous normal squamous epithelium tissues. Expression was statistically analyzed with regard to clinicopathological factors using chi(2) and non-parametric tests. The expression level of DRP-1 was significantly different between the ESCC and paracancerous tissues (P<0.05). The expression level was correlated with the depth of invasion and lymph node metastasis (P<0.05). Abnormal E-cadherin expression was found to be associated with a high degree of cancer differentiation and lymph node metastasis (P<0.05). A positive correlation was observed between the expression of DRP-1 and E-cadherin (P<0.05). The expression of ezrin was found to be correlated with the depth of ESCC invasion, the degree of differentiation and lymph node metastasis (P<0.05). The high expression of ezrin has been previously shown to be correlated with the low or absent expression of E-cadherin. In conclusion, in ESCC, the expression levels of DRP-1, ezrin and E-cadherin were all reduced, and this reduction or absence of expression may have been attributed to ESCC tumorigenesis and progression. Simultaneous analyses of DRP-1, ezrin and E-cadherin expression levels would be useful to determine the malignancy and metastatic potential of ESCC, and these genes are consequently of potential use as biomarkers for the diagnosis and prognosis assessment of early-stage ESCC.