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Background Twist is a highly conserved epithelial-mesenchymal transcription factor that has been reported to be a key factor in tumor malignancy, including lymph node metastasis. It represents the major step of dissemination and serves as a chief prognostic indicator of disease progression. However, the mechanism by which Twist regulates lymph node metastasis remains incompletely understood. Studies on the mechanism of metastasis are thus required for determining appropriate therapeutic strategies.Methods Immunohistochemistry for lymphatic vessel endothelial receptor 1 (LYVE-1), Ki-67, Twist, vascular endothelial growth factor C (VEGF-C), and vascular endothelial growth factor receptor 3 (VEGFR-3) was performed to detect lymphatic vessel density (LVD), cell proliferation levels and the expressions of Twist, VEGF-C, and VEGFR-3 were determined from 66 primary supraglottic carcinoma tissue samples from 36 patients with lymph node metastasis (pathological N+, pN+) and 30 patients without metastasis (pathological NO, pNO). Western blotting analysis of the proteins in pN+ and pNO primary tumors was used to characterize the expressions of Twist, VEGF-C, and VEGFR-3further.Results The LVD was 22.4±10.3 in pN+ patients and 6.8±4.1 in pNO ones. For Ki-67, the number of proliferous cells in pN+ patients was greater than that in pNO ones. Both, however, were associated with their clinical nodal stages. In pN+patients, Twist, VEGF-C, and VEGFR-3 expressions were 86.11% (31/36), 80.56% (29/36), and 58.33% (21/36),respectively. These values were higher than those found for pNO patients (i.e., 13/30, 11/30, and 7/30, respectively) (P ＜0.05). Among the samples with Twist expression, 88.64% were VEGF-C-positive and 59.09% were VEGFR-3-positive.The pNO counterparts were 4.55% and 9.09%, respectively (P＜0.05). The expressions of Twist, VEGF-C, and VEGFR-3in pN+ patients obtained through Western blotting analysis were significantly higher than those in pNO patients, and the levels of VEGF-C and VEGFR-3 were positively correlated with that of Twist.Conclusions Twist expression correlates with lymph node metastasis. The mechanism involved in such a correlation may be related to lymphangiogenesis.