Matrilysin (MMP-7) is over-expressed in various cancers and is thought to play important roles in tumor invasion and metastasis. However, the function of MMP-7 in breast cancer remains unclear. We therefore examined the expression of the MMP-7 gene in breast cancer (MCF-7) cells and the effect of its over-expression on cellular invasion. We transfected human MMP-7 into MCF-7 cells and selected clones that stably over-expressed the MMP-7 gene. The in vitro invasiveness of MCF-7 cells was quantified by use of the Matrigel invasion assay. Expression of MMP-7 mRNA was analyzed by quantitative RT-PCR. MMP secretion and activation were detected by gelatin zymography. We found that MMP-7-expressing clones had significantly increased invasion (P < 0.001), with increased MMP-7 expression and gelatinase activation as compared to the vector controls. We conclude that MMP-7 over-expression correlates with breast cancer in vitro invasiveness and that MMP-7 may promote invasion by increasing the secretion and activation of proMMP-2 and proMMP-9.