Abstract: Objective: To intensively investigate the clinicopathologic significance of the alternadons of p16 gene in esophageal squamous cell carcinoma (ESCC), so as to provide a reliable indicator and create a new way for the research on clinical prevention and treatment of ESCC. Methods : Sixtynine samples from the patients with ESCC were enrolled. PCR-SSCP and DNAsequencing were utilized to detect the mutation and deletion of p16 gene exonl and exon2. Results : Thirty-three cases with p16 gene abnormality including sixteen point mutations and seventeen deletions, which were defined as Group M, were detected in sixty-nine cases of No p16 gene abnormality was found in the rest of thirty-six cases, which were classified as GroupN. There were significant differences in lymph node metastasis and distance metastasis between these groups (P<0.05). However, no significant differences in sex,age, the length and invasion’s depth of the tumor (P>0.05) were found. Conclusions : The mutations of p16 gene may be the late changes in ESCC. Detecting the mutation of p16 gene may serve as a marker of predicting degree of maligmancy, development and prognosis. It will render an authentic evidence for the selection of the clinical cases with high risk of metastasis.