In order to establish a MCF-7 cell line with lentivirus-mediated FOXP2 gene silencing and confirm its interference effect, the lentiviral vector (pMAGic7.1-shFOXP2) was constructed by genetic engineering and cotransfected with pVSVG、Δ8.91 into HEK293T cells, which generated the mature lentivirus, viral supernatant was collected and added to MCF-7 cells after 48 h, the mRNA and protein levels of FOXP2 were examined by qPCR and Western blot. Thus, we constructed and identified the lentiviral recombinant vector pMAGic7.1-FOXP2,successfully established a MCF-7 cell line with the interference of FOXP2 expression, as these findings imply a promising strategy for studying FOXP2 functions in breast cancer progression.