Colorectal cancer (CRC) is one of most common cancers and causes of cancer-associated mortality worldwide, due to its recurrence, metastasis and therapy resistance. Cancer stem cells (CSC) have been demonstrated to be vital for tumor initiation and recurrence. microRNAs may act as an oncogenes or tumor suppressors in numerous cancers. The present study demonstrated that microRNA-141 (miR?141) was downregulated in CSC compared with differentiated cancer cells, and in tumor compared with healthy tissue. miR?141 may inhibit CRC cell proliferation and the maintenance of CSC stemness, thereby enhancing drug susceptibility. In addition, the present study identified cyclin D2 as a novel target gene of miR?141. In conclusion, the antitumor role of miR?141 and its target cyclin?D2 may suggest the development of miR?141 as a potential therapeutic agent.