General Background: Breast cancer is the most prevalent cancer affecting women, with increasing incidence worldwide. Specific Background: Recent research has focused on the role of epigenetic changes in DNA damage, repair mechanisms, and the potential therapeutic effects of probiotics. Probiotics have shown promise in promoting tissue regeneration and DNA repair. Knowledge Gap: However, the precise impact of probiotics on DNA repair in cancer cells, specifically breast cancer cells, remains underexplored. Aims: This study aimed to evaluate the effects of probiotics on DNA damage repair in AMJ13 Iraqi breast cancer cells and assess the cytotoxic effects of probiotics on these cells. Results: Using the comet assay, we found significant increases in DNA damage repair in AMJ13 cells treated with Lactobacillus plantarum (T1) and a combination of eight probiotic strains (T2). Exposure to T1 for 48 hours resulted in significant increases in tail DNA (P≤0.001), head DNA (P≤0.001), and tail moment (P<0.001), while T2 showed similar significant increases at 72 hours (P<0.05). Image analysis further supported the DNA repair potential of probiotics, as indicated by a small tail curve for treated cells. Novelty: This study provides novel insights into the therapeutic potential of probiotics in breast cancer treatment by demonstrating their capacity to enhance DNA repair mechanisms in cancer cells. Implications: The findings suggest that probiotic therapy may be a promising adjunct treatment in breast cancer, offering a new avenue for cancer management through the enhancement of DNA repair and reduction of DNA damage. Highlights:   Probiotics significantly repaired DNA damage in breast cancer cells. T1 and T2 enhanced DNA repair within 48-72 hours. Probiotics offer potential as breast cancer adjunct therapy.   Keywords: Breast cancer, probiotics, DNA repair, AMJ13 cells, cytotoxicity