The microRNA (miR/miRNA)-182/183/96 cluster comprises miR-96, -182 and -183. The present study examined five previous microarray-based human colon cancer miR expression profiling studies and the expression of these three miRs was found to be upregulated in colon cancer tissues. Subsequently, in vitro assays were performed to determine the role of the miR-183/182/96 cluster in colon cancer cells. The results demonstrated that inhibiting miR-183, miR-182 or miR-96 with antisense oligonucleotide (ASO)-mimics inhibited the proliferation of colon cancer cells. Notably, further investigation revealed that inhibiting their expression simultaneously led to a more efficient reduction in cancer cell proliferation. These results suggested that miR-182/183/96, which resides in clusters in the genome, functioned synergistically in colon cancer and implied that co-expression of the miR cluster ASOs was efficient in reducing tumorigenesis, offering novel insight into the use of miRNAs in tumor therapy.