MicroRNAs play critical roles in the development and progression of colorectal cancer (CRC). miR-154 acts as a tumor suppressor in several tumors; however, its role in CRC is poorly understood. Herein, we found that miR-154 was decreased in CRC tissues and cell lines. Ectopic expression of miR-154 remarkably suppressed cell proliferation and colony formation, migration and invasion in CRC cells. The toll-like receptor 2 (TLR2) was found to be a direct target of miR-154 in CRC cells. Inhibition of TLR2 performed similar effects with miR-154 overexpression on CRC cells, and overexpression of TLR2 could significantly reverse the tumor suppressive effects of miR-154 on CRC cells. This study suggests an essential role for miR-154 in CRC.