To determine the expression and function of B cell translocation gene 1 (BTG1) in nasopharyngeal carcinoma. Nasopharyngeal samples were taken from cancer lesions (n = 75) and adjacent normal tissue (n = 33) in nasopharyngeal cancer patients immediately after endoscopic biopsy. BTG1 expression was determined by immunohistochemistry and Western blotting. The effect of BTG1 overexpression was examined in vitro utilizing a human nasopharyngeal cancer cell line CNE2 stably transfected with a recombinant lentivirus (LeBTG1 cells) and compared to empty vector-transfected controls (LeEmpty). BTG1 overexpression was verified by real-time reverse transcriptase polymerase chain reaction and Western blot. The expression of proteins involved in cell cycle regulation (cyclin D1), apoptosis (Bcl-2) and cell migration (MMP-9) in LeBTG1 cells were analyzed by Western blot. The effect of BTG1 overexpression on cell viability and proliferation was assessed by an MTT assay in LeBTG1 and LeEmpty cells. Flow cytometric analyses were used to evaluate the effect of BTG1 expression on cell cycle distribution and apoptosis. The migration and invasion potential of LeBTG1 cells was examined by plating cells in Matrigel-coated chambers. BTG1 protein expression was significantly lower in nasopharyngeal cancer tissue biopsies than normal tissue as measured by immunohistochemistry (36.0 vs. 81.8 % of tissues; P < 0.05) and Western blotting (0.221 +/- A 0.019 vs. 0.652 +/- A 0.055; P < 0.05). Decreased expression of BTG1 was significantly correlated with nasopharyngeal cancer tumor stage, lymph node metastasis, clinical stage and pathologic differentiation (P < 0.05), as well as with reduced overall five-year survival rates compared to patients with higher expression levels (31.2 vs. 70.2 %; P < 0.05). In vitro analyses revealed that LeBTG1 cells had a reduced survival fraction compared to control LeEmpty cells, with higher rates of apoptosis (9.3 +/- A 0.7 vs. 2.3 +/- A 0.3 %; P < 0.05). The proportion of LeBTG1 cells in G0/G1 stage and S phase was also significantly different from LeEmpty cells (82.6 +/- A 3.8 and 10.1 +/- A 1.0 %, vs. 62.2 +/- A 2.4 and 28.9 +/- A 2.0 %, respectively; Ps < 0.05), and the migration and invasion of LeBTG1 cells was significantly impaired with respect to LeEmpty cells (96.0 +/- A 13.0 and 91.0 +/- A 11.0 vs. 158.0 +/- A 17.0 and 142.0 +/- A 15.0, respectively; Ps < 0.05). These effects were accompanied by decreased protein expression of cyclin D1, Bcl-2 and MMP-9 in LeBTG1 cells (0.231 +/- A 0.021, 0.413 +/- A 0.046, 0.131 +/- A 0.011, respectively) compared to control LeEmpty cells (0.636 +/- A 0.067, 0.821 +/- A 0.083, 0.451 +/- A 0.041, respectively; Ps < 0.05). Reduced BTG1 expression is associated with increased disease severity, suggesting it is a negative regulator of nasopharyngeal cancer and can serve as a prognostic indicator.