Abnormal microRNA expression is a common and important feature of human malignancies. Matrix metalloproteinase 2 (MMP2), which has been reported in several cancers, plays important roles in cancer progression. However, the microRNA regulatory mechanism on MMP2 expression remains unclear. In this study, we first detected MMP2 and microRNA-29a (miR-29a) expression in oral squamous carcinoma (OSCC) specimens, which showed that MMP2 was higher in OSCC cancer tissues than adjacent tissues but that miR-29a was lower in OSCC cancer tissues than adjacent tissues. Then, we confirmed that miR-29a, which directly targeted 3′-UTR of MMP2 gene, negatively regulated MMP2 expression by miR-29a transfection and luciferase reporter assay. Exogenous overexpression of miR-29a inhibited OSCC cell invasion and anti-apoptosis significantly in vitro. Whereas, knockdown of miR-29a promoted OSCC cell invasion and induced drug-resistance in vitro. This study suggests that miR-29a plays an inhibiting role in the progression of OSCC, which may be a potentially therapeutic approach in the future.