BI-1 (Bax inhibitor-1), an apoptosis-inhibiting gene belonging to the Bcl-2 protein family, plays an important role in mitochondrial apoptosis pathway to suppress Bax-induced apoptosis. To investigate the potential role of BI-1 in promoting cell growth and tumorigenesis, in the present study we overexpressed the BI-1 gene in NIH3T3 cells using the lentivirus-mediated gene expression system. Our in vitro studies showed that NIH3T3 cells overexpressing BI-1 displayed a significantly higher growth rate and formed more and larger colonies than the control cells. In addition, our in vivo studies indicated that the lenti-BI-1-infected cells formed obvious tumours, while no tumours were formed by the control cells after subcutaneously injected into nude mice. These results strongly suggested that the BI-1 gene might play a crucial role in neoplastic genesis and development.