Breast cancer is one of the most common malignancies in women. ERα, PR, and HER2 triple nega-tive breast cancer (TNBC) is the current research focus because of the lack of effective targetedtherapies. In our study, lentivirus systems were used to overexpress EZH2 and miR-1301 in TNBCcell lines. Western blot analysis and RT-qPCR were used to detect the protein and microRNAlevels. The TCGA and Kaplan Meier plotter databases were used to analyze the EZH2 and miR-1301 expression levels in breast cancer. The effect of miR-1301 overexpression on cell prolifer-ation, migration and colony formation were determined by using the sulforhodamine B (SRB)assay, wound healing assay and colony formation assay, respectively. Furthermore, an xenograftmouse model was used to investigate the function of miR-1301 overexpression in vivo. Finally,dual luciferase reporter assay was used to verify the binding site of EZH2 and miR-1301. We foundthat EZH2 induced the expression of miR-1301 in two TNBC cell lines, HCC1937 and HCC1806.Overexpression of miR-1301 suppressed TNBC cell proliferation, migration and colony formation,as well as the xenograft tumor growth in immunodeficient mice. Interestingly, miR-1301 inhibitedthe expression of EZH2 by binding to the 3′-UTR of EZH2 gene. These data suggest that EZH2induces the expression of miR-1301 as a negative feedback control mechanism in TNBC.