In Vivo Investigation of Supportive Immunotherapeutic Combination of Bifidobacterium infantis 35624 and Doxorubicin in Murine Breast Cancer.

The aim of the study is to investigate the anti-tumor effect of Bifidobacterium infantis 35624 in a xenograft model in BALB/c mice injected with 4T1 cells as a support for chemotherapeutic treatments of doxorubicin in vivo. The MTT assay was used to determine the cytotoxicity of doxorubicin against cancer cells, and apoptosis was analyzed by using flow cytometry. 4T1 cells (2 × 104 cells/mouse) were injected to BALB/c mice, and mice were fed with/without gavage B. infantis milk (108 CFU/mL) for 14 days and treated with doxorubicin on 5th and 10th days. The weights of the mice were recorded during the study, and the tumor sizes were measured by caliper at the 14th day. CD8 + T cell response was analyzed by using flow cytometer, and the results were compared to control and tumor control groups. The IC50 value for doxorubicin on 4T1 cell lines was determined as 0.053 ± 0.012 µg/mL. The apoptotic effect of doxorubicin at IC50 concentration was determined as 82.3% of cells to late apoptosis, 3.6% of cells to pro-apoptosis, and 6.2% of cells to necrosis. The treatment of doxorubicin, B. infantis milk, and the combination of them inhibited the tumor volumes by 55.50%, 40.69%, and 75.95%, respectively. B. infantis administration significantly enhanced the PHA-induced splenocyte proliferation (P < 0.05). It was shown that IFN-γ was effective in tumor growth and regression of metastasis. Consequently, the combination of B. infantis milk and doxorubicin showed the best anti-tumor effect.