Objective To investigate the effect of microRNA-584-5p (miR-584-5p) on the biological behavior (proliferation, migration and invasion) of breast cancer cells and its mechanism. Methods Human normal breast epithelial cells MCF10A and breast cancer cells MDA-MB-231, SK-BR-3 and MCF-7 were selected; take MCF-7 cells in logarithmic growth phase, transfect them with LipofectamineTM 2000 transfection kit, and divide them into seven groups: blank group (untransfected MCF-7 cells), mimic-negative control (mimic-NC) group (transfected mimic-NC), miR-584-5p mimic group (transfected miR-584-5p mimic), pcDNA group [transfected with overexpression of matrix metalloproteinase-14 (MMP-14) pcDNA3.1 plasmid negative control (pcDNA3.1)], MMP-14 group [transfected with overexpression of MMP-14 pcDNA3.1 plasmid (pcDNA3.1-MMP-14)], mimic-NC+MMP-14 group (co-transfected with mimic NC and pcDNA3.1-MMP-14), and miR-584-5p mimic+MMP-14 group (co-transfected with miR-584-5p mimic and pcDNA3.1-MMP-14). The mRNA expression levels of miR-584-5p in MCF10A, MDA-MB-231, SK-BR-3 and MCF-7 cells and the expression levels of miR-584-5p and MMP-14 mRNA of MCF-7 cell in each group were detected by fluorescence quantitative PCR. The protein expressions of MMP-14 of MCF-7 cell in each group were detected by Western blotting. The proliferation, migration and invasion of MCF-7 cell in each group were detected by cell counting kit - 8 (CCK-8), scratch test and Transwell test. The targeting relationship between miR-584-5p and MMP-14 was detected by double luciferase reporter gene assay. Results Compared with the human normal mammary epithelial cells MCF10A, the expression levels of miR-584-5p in breast cancer cells MDA-MB-231, SK-BR-3 and MCF-7 were decreased (P<0.05), and the expression level of miR-584-5p in MCF-7 cells was the lowest. Compared with the blank group and the mimic-NC group, the expression level of miR-584-5p of MCF-7 cells in the miR-584-5p mimic group was increased, and the expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number were decreased or reduced (P<0.05). Compared with the blank group and the pcDNA group, the expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number of MCF-7 cells in the MMP-14 group were increased (P<0.05). Compared with the MMP-14 group and the mimic-NC+MMP-14 group, the expression level of miR-584-5p of MCF-7 cells in the miR-584-5p mimic+MMP-14 group was increased, the expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number were decreased or reduced (P<0.05). The expression levels of MMP-14 mRNA and protein, proliferation activity, scratch healing rate and invasive cell number of MCF-7 cells in the miR-584-5p mimic+MMP-14 group were higher or morer than those in the miR-584-5p mimic group (P<0.05). The results of double luciferase reporter gene test showed that miR-584-5p could targeted action on the MMP-14 promoter region. Conclusions MiR-584-5p can targetable regulate the expression of MMP-14. Overexpression of miR-584-5p inhibits the proliferation, migration and invasion of breast cancer cells by down-regulating MMP-14.