Among seven strains of lactic acid bacteria (LAB) isolated from traditional dairy products, a Lactobacillus strain was identified through 16S rRNA gene sequencing and tentatively designated as Lactobacillus brevis MK05. This strain demonstrated the highest probiotic potential through biochemical analysis, including acid and bile salt resistance, as well as antibacterial activity. The collected cell-free supernatant (CFC) of L. brevis MK05 culture, compared with MRS broth with pH equal to the pH for CFC, revealed antimicrobial activity against Escherichia coli (ATCC 25922) and Staphylococcus aureus subsp. aureus (ATCC 25923), possibly due to the presence of antibacterial metabolites other than organic acids. This strain was, therefore, selected to assess the biological activity of its partially purified secretory proteins against MCF-7 cancer cells and normal fibroblast cells via the MTT assay. The partially purified cell-secreted proteins of this strain (hereafter referred to as Lb-PPSPs) showed a time and dose-dependent anti-cancer and apoptosis induction function. There was a remarkable decline in the survival rate of MCF-7 cells at doses equal to and higher than 0.5 mg/mL after 48 h. The changes in expression of the three genes involved in the apoptosis pathway (BAX, BCL-2, and BCL2L11) in MCF-7 cells treated with the Lb-PPSPs confirm its cytotoxic activity and apoptosis induction.