This study aimed to assess the anti-proliferative capabilities of the three probiotic strains on breast cancer (MCF7) and test their anticancer capabilities onRP1 and RASSF1A Genes. Three probiotics bacterial strains: Lactobacillus casei ss. casei (LC 1093), Lactobacillus delbreuckii ss. bulgaricus LD (1102) and Bifidobacterium bifidum (BB 1334) were tested for their anti-proliferative capabilities on cell lines via trypan blue test and MTT assay. Their anti-methylation activities were tested using methylation-specific PCR (MSP). Results revealed that Lactobacillus casei strain achieved the highest percentage of cancer cell death. The effects of these strains on the methylation status of RASSF1A and RB1 promotor regions in breast cancer cells were tested. The unmethylation-specific primers of both genes were able to generate a defined band. The methylation patterns were reshaped when compared to the untreated MCF7 cell line showing the epigenetic delaying mechanism of the probiotic cell free filtrate by interfering the methylation mechanism of breast cancer on two tested genes.