Dysregulated expression of microRNAs (miRNA) is a hallmark of cancer. miR-16 has been reported to be downregulated and to act as a tumor suppressor in different cancer types. In the present study, we sought to investigate the possible roles and mechanisms of miR-16 and its relationship with p53 and survivin in CRC cells. We showed that miR-16 was downregulated in 67% of CRC tissues and was correlated with the degree of histological differentiation. Experiments in vitro showed that overexpression of miR-16 inhibited the proliferation and induced apoptosis of CRC cells through the intrinsic apoptosis pathway. We further showed that miR-16 repressed survivin expression at both the mRNA and protein levels and the survivin gene was a direct target of miR-16. In addition, miR-16 reduced p53 expression and p53 increased miR-16 levels, with downregulation of miR-16 targets survivin, cyclin D1 and CDK6. Our findings suggest that miR-16 represses colorectal cancer cell growth in vitro by regulating the p53/survivin signaling pathway. Our findings provide further evidence for the involvement of dysregulated miRNAs in CRC, and miR-16 could serve as a molecular target for CRC therapy.