We determined the expression and function of B cell translocation gene 1 (BTG1) in thyroid carcinoma. Thyroid samples were obtained from cancer lesions (n=83) and adjacent normal tissue (n=35) in thyroid 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 the human thyroid cancer cell line FTC-133, stably transfected with a recombinant lentivirus (LeBTG1 cells) and compared to empty vector transfected controls (Le Empty). BTG1 overexpression was verified by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting. The expression of proteins involved in cell cycle regulation (cyclin D1), apoptosis (Bcl-2) and cell migration (MMP-9) in LeBTG1 cells was analyzed by western blotting. The effect of BTG1 overexpression on cell viability and proliferation was assessed by MTT assay in LeBTG1 and Le Empty 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 thyroid cancer tissue biopsies compared to normal tissue as measured by immunohistochemistry (36.1 vs. 80.0% of tissues; P<0.05) and western blotting (0.251 +/- 0.021 vs. 0.651 +/- 0.065; P<0.05). Decreased expression of BTG1 was significantly correlated with thyroid cancer lymph node metastasis, clinical stage and pathological differentiation (P<0.05), as well as with reduced overall 10-year survival rates compared to patients with higher expression levels (30.2 vs. 66.7%; P<0.05). In vitro analyses revealed that LeBTG1 cells had a reduced survival fraction compared to control Le Empty cells, with higher rates of apoptosis (11.6 +/- 2.1 vs. 2.1 +/- 0.4%; P<0.05). The proportion of LeBTG1 cells in G0/G1 stage and S phase was also significantly different from Le Empty cells (81.8 +/- 6.3 and 10.2 +/- 1.0%, vs. 62.4 +/- 4.9 and 25.5 +/- 2.6%, respectively; P<0.05), and the migration and invasion of LeBTG1 cells was significantly impaired with respect to Le Empty cells (72.0 +/- 8.0 and 55.0 +/- 7.0 vs. 113.0 +/- 16.0 and 89.0 +/- 9.0, respectively; P<0.05). These effects were accompanied by decreased protein expression of cyclin D1, Bcl-2 and MMP-9 in LeBTG1 cells (0.234 +/- 0.018, 0.209 +/- 0.021, 0.155 +/- 0.017, respectively) compared to control Le Empty cells (0.551 +/- 0.065, 0.452 +/- 0.043, 0.609 +/- 0.072, respectively; P<0.05). Reduced BTG1 expression is associated with increased disease severity, suggesting it is a negative regulator of thyroid cancer and can serve as a prognostic indicator.