MicroRNAs (miRNAs) are a class of small non‑coding RNA molecules, which play regulatory roles at the post-transcriptional level by suppressing the translation of protein‑coding genes or inducing mRNA cleavage. Dysregulated expression of miRNAs is involved in multiple types of cancers and plays important roles in regulating various biological processes including metastasis. miR-101 is downregulated in various types of cancer and functions as a suppressor of cell migration and invasion. Meanwhile, enhancer of zeste homolog 2 (EZH2) is associated with the metastatic potential of several aggressive tumors. In the present study, we reported that ectopic overexpression of miR-101 downregulated the expression level of EZH2 and significantly inhibited migration and invasion of osteosarcoma cells. In addition, knockdown of EZH2 by siRNA showed the same effect of miR-101 on migration and invasion. To conclude, these results indicate that miR-101 may act as a tumor suppressor in osteosarcoma, as it has a suppressive role in cell migration and invasion by targeting EZH2.