Objective: To evaluate the expression and correlation of hypoxia inducible factor 1α (HIF-1α) and vascular endothelial growth factor (VEGF) and Survivin proteins in biopsy specimens of esophageal squamous cell carcinoma (ESCC), and then determine whether the levels of expression of these proteins could predict the clinical effectiveness of radiotherapy in individual cancers. Methods: The expressions of HIF-1α, VEGF and Survivin were shown by S-P immunohistochemical staining method in biopsy specimens of ESCC, which were obtained endoscopically from 50 patients before radiotherapy, and 10 cases of normal esophageal tissue. Results: The positive expression rates of HIF-1α, VEGF and Survivin were 68%, 74% and 72% in ESCC respectively. However, the three tumor markers had negative expressions in normal esophageal tissue. The positive rate of HIF-1α was positively correlated with VEGF and Survivin proteins. The positive rates of HIF-1α and Survivin were closely related to the clinical stage, radiotherapy effectiveness and survival, otherwise, the expression of HIF-1α was closely related to distant metastasis; both of them were no correlation with the differentiation degree of tumor. The effective rates of radiotherapy and mean survival periods of those cases with positive and negative expressions of HIF-1α were 8.8%, 10 months and 81.25%, 25 months, respectively. The one, two, and three years survival rates of patients with positive and negative expressions of HIF-1α were 38.2%, 5.9%, 2.9%, and 81.3%, 54.2%, 15.8%, respectively (P = 0.001). Patients with HIF-1α positive expression obviously survived less than those with negative expression and the difference was significant. The expression of VEGF was only related to the distant metastasis. Conclusion: Over expressions of HIF-1α, VEGF and Survivin were found in ESCC. The positive rate of HIF-1α was positively correlated with VEGF and Survivin proteins. The expression of HIF-1α may serve as an important parameter in evaluating response for radiotherapy and prognosis of ESCC. It may play an important role by up-regulating the transcription of VEGF and Survivin genes.