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DDX 1, a member of the  DEAD  box  RNA  helicase family, plays a critical role in testicular tumors. However, it remains to be clarified whether  DDX 1 is involved in other types of malignant tumors such as colorectal cancer. We disrupted the  DDX1  gene in a human colorectal cancer cell line LoVo using the  CRISPR /Cas9‐mediated gene‐targeting system.  DDX 1‐ KO  LoVo cells exhibited a much slower growth rate, produced fewer colonies in soft agar medium, and generated smaller solid tumors in nude mice than parental LoVo cells. Such phenotypes of the  DDX 1‐ KO  cells were mostly reversed by exogenous expression of  DDX 1. These results indicate that  DDX 1 is required for tumorigenicity of colorectal cancer cells. In the  DDX 1‐ KO  cells, the cancer stem cell marker genes  LGR5 ,  CD133 ,  ALDH1  and  SOX2  were markedly suppressed. Among them, expression of  LGR 5, which is essential for tumorigenicity of colorectal cancer cells, was restored in the  DDX 1‐transfected  DDX 1‐ KO  cells. Consistently, the  DDX 1‐ KO  cells lost sphere‐forming capacity in a  DDX 1‐dependent fashion. Reporter and chromatin immunoprecipitation assays revealed that  DDX 1 directly bound to the −1837 to −1662 region of the enhancer/promoter region of the human  LGR5  gene and enhanced its transcription in LoVo cells. Repression of  LGR5  by  DDX 1 knockdown was observed in 2 other human colorectal cancer cell lines, Colo320 and  SW 837. These results suggest that  LGR 5 is a critical effector of  DDX 1 in colorectal cancer cells. The  DDX 1‐ LGR 5 axis could be a new drug target for this type of malignant cancer.

DEAD box protein DDX 1 promotes colorectal tumorigenesis through transcriptional activation of the LGR5 gene