Numerous microRNAs (miRNAs) are dysregulated in tongue squamous cell carcinoma (TSCC), and their dysregulation has been demonstrated to have a strong correlation with TSCC progression via regulation of their targets. Therefore, miRNAs have potential use in the diagnosis and treatment of patients with TSCC. In the present study, miRNA-758 (miR-758) expression in TSCC tissues and cell lines was detected through reverse transcription-quantitative polymerase chain reaction, and the effects of miR-758 on TSCC cell proliferation and invasion were investigated by using Cell Counting kit-8 and Transwell invasion assays. A luciferase reporter assay was performed to determine the target interaction between miR-758 and metadherin (MTDH) in TSCC cells. The results revealed that miR-758 was downregulated in TSCC tissues and cell lines. miR-758 overexpression restricted the proliferation and invasion of TSCC cells. Additionally, MTDH was verified as a direct target gene of miR-758 in TSCC cells. Furthermore, MTDH was observed to be upregulated in TSCC tissues, and the upregulation of MTDH was inversely correlated with miR-758 expression. Moreover, restored MTDH expression significantly counteracted the suppressive effects of miR-758 overexpression on TSCC cells. These results suggested that miR-758 may prevent TSCC progression and development by directly targeting MTDH, thereby providing evidence that miR-758 is a novel therapeutic target for the treatment of patients with TSCC.