Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in southern China and Southeast Asia. It is characterized as a multistep process involved in multiple genetic and epigenetic events. The mechanism of carcinogenesis still needs to be further clarified. In this study, two-dimensional gel electrophoresis, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), RT-PCR, Western blot and immunohistochemical (IHC) analyses were used to detect Galectin-1 expression in NPC compared with normal nasopharyngeal epithelial tissues (NNET). We found that Galectin-1 was expressed at a significantly higher level in NPC compared with NNET. Our results indicated that high expression level of Galectin-1 might correlate with the development of NPC and Galectin-1 may serve as a potential diagnostic marker or therapeutic target for NPC.