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Background:The purpose of the present investigation is to unravel the influence of the miR-375 inhibitor on cell survival descent and apoptosis stimulation, mediating PI3K/Akt/mTOR signaling network in human breast cancer cells. Methods: MCF-7 cells were transfected with miR-375 inhibitor for 72 h. Then, cell survival assay was measured by MTT. Cells were stained with EtBr/AO, DAPI, and DCFH-DA to assess the effect of the miR-375 inhibitor on cell death. A scratch experiment was performed to observe the cell migration ability. The expression of anti-apoptotic and apoptotic genes such as BCL-2, BAX, and PI3K/Akt/mTOR and miR-375 were evaluated in miR-375 inhibitor transfected cells by qRT-PCR. Findings: MiR-375 inhibitor sensitized tumor cells and influenced significant loss in the breast cancer cell proliferation with obvious cell death elevation. MiR-375 inhibitor effectively augmented ROS generation. Also, miR-375 inhibition hampered migratory ability. Furthermore, our qRT-PCR analysis showed that inhibition of the miR-375 was able to significantly reduce the constitutive expression of PI3K/Akt/mTOR mRNAs. Additionally, miR-375 suppression decreased the anti-apoptotic gene, Bcl-2 expression and enhanced pro-apoptotic gene, Bax expression along with potentially decreasing miR-375 level compared to control. Novelty and applications: Inhibition of the miR-375 has considerably attenuated cell proliferation and stimulated apoptotic cell death in the breast cancer cells. Thus, miR-375 represent a potential therapeutic target for the breast cancer.