Introduction: Lactic acid bacteria, recognized as probiotics, have garnered significant attention as potential adjuvants in chemotherapy for various cancer types, including cervical cancer. In this study, we investigated the anti-cancer properties of two indigenous Iranian strains, Lactobacillus acidophilus and Lactobacillus paracasei, individually and in combination, targeting human cervical cancer cell lines compared to normal control cells. Methods: The cytotoxic effect of Lactobacillus acidophilus and Lactobacillus paracasei supernatants, as well as their 1:1 mixture, on CaSki and HNCF PI 52 cell lines, was evaluated using the MTT assay. The apoptotic and anti-metastatic effects of these supernatants were assessed by analyzing the gene expression of BAX/BCL2 ratio, Caspase-3, and MMP2/ MMP9 using Real-Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Results: Significant cytotoxicity was observed in Ca Ski cells attributed to the low pH of the supernatants. The increase in the BAX/BCL2 ratio, leading to an up-regulation of Caspase-3, indicated the induction of apoptosis (P<0.001). In addition, the expression of MMP9 significantly deceased in Ca Ski cells treated with Lactobacillus acidophilus (P<0.001) and Lactobacillus paracasei (P<0.05), while no significant difference in MMP2 expression was observed in all samples compared to the control groups. Conclusion: while further validation is needed, the heightened expression of apoptotic genes suggests a potential induction of apoptosis in cancer cells in response to Lactobacillus toxicity. The significant down-regulation of the MMP9 gene emphasizes the need for comparative analyses across different cervical cancer cell lines to establish the anti-metastatic potential of these local probiotic supernatants.