Nuclear receptor binding protein 1 (NRBP1) is an evolutionarily highly conserved adaptor protein with multiple domains. Recently, its role in cancers has received increasing attention. To investigate whether NRBP1 is involved in the development of bladder cancer, we used tissue microarray and analyzed the association between the expression levels of NRBP1 and clinico-pathological features of 56 patients diagnosed with bladder cancer. Subsequently, NRBP1 was silenced using siRNA in bladder cancer cell lines T24 and 5637, and cell phenotype such as proliferation and apoptosis were observed. Further, in vivo tumor formation assay was performed. The expression of apoptosis markers was detected by Western blot. A significant positive correlation between increased NRBP1 expression and tumor stage, and lymph node metastasis was observed in 56 patients. High expression of NRBP1 was associated with poor prognosis and NRBP1 knockdown significantly inhibited cell proliferation and induced intrinsic apoptosis in vitro. Moreover, we also found that NRBP1 knockdown significantly suppress tumor growth in xenograft mouse model. Taken together, these data suggest that NRBP1 plays a critical role in the development of bladder cancer and may represent a potential target for bladder cancer treatment.