The targeted delivery of small interfering RNA (siRNA) to specific tumor tissues and tumor cells remains as one of the key challenges in the development of RNA interference as a therapeutic application. The ribosome assembly factor NIN/RPN12 binding protein (NOB1) has been suggested to be essential for processing of the 20S pre-rRNA to the mature 18S rRNA, and is also reported to participate in proteasome biogenesis. However, it is unclear whether NOB1 is involved in tumor cells growth. The aim of this study was to determine whether the suppression of lentivirus mediated NOB1 siRNA inhibits the growth of human clean cell carcinoma (ccRCC) cells, further focused on NOB1 as a possible therapeutic target for renal cell carcinoma treatment. NOB1 deletion that caused significant decline in cell proliferation was observed in both 786-O and ACHN cell lines as investigated by MTT assay. Further, the number and size of the colonies formed were also significantly reduced in the absence of NOB1. Moreover, NOB1 gene knockdown arrested the cell cycle and inhibited cell cycle-related protein expression. The Kaplan-Meier survival curves revealed that low NOB1 expression was associated with poor prognosis in ccRCC patients. Collectively, these results indicate that NOB1 plays an essential role in renal cell cancer cell proliferation, and its gene expression could be a therapeutic target.