MicroRNA (miRNA, miR) is a small endogenous noncoding RNA and is involved in the pathogenesis of a variety of human cancers. However, the potential roles of miR-127-3p in mucoepidermoid carcinoma (MEC) have not been studied. The purpose of this study is to identify the role of miR-127-3p in MC-3 human MEC cells. The growth inhibitory effect and related mechanism of miR-127-3p was performed using MTS assay, Flow cytometry analysis, DAPI staining, anchorage-independent cell transformation assay and Western blot analysis. The transfection of exogenous miR-127-3p into MC-3 cells decreased cell viability, and it caused the G1/S cell cycle arrest. MiR-127-3p also decreased neoplastic cell transformation in TPA-induced JB6 mouse epidermal cells and MC-3 cells. In addition, miR-127-3p decreased specificity protein 1 (Sp1) expression and increased p21 and p27 expression which are Sp1-dependent cell cycle-related proteins. However, miR-127-3p did not induce apoptosis or affect myeloid cell leukemia-1 (Mcl-1) and survivin. MiR-127-3p induced G1/S cell cycle arrest and increased p21 and p27 expression via the modulation of Sp1. Thus, miR-127-3p might be a potential therapeutic target for human mucoepidermoid carcinoma.