Dendritic cells-based vaccine to inhibit triple-negative breast cancer cells proliferation.
ABSTRACT
Background: Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer with limited effective treatment options. New therapeutic approaches are urgently needed to improve the prognosis of TNBC. We studied the character of Runx2 gene in breast cancer and whether the Runx2-dendritic cell (DC) vaccine can induce specific antitumor effect on triple negative breast cancer in vitro.
Methods: The expression of Runx2 in triple negative breast cancer，non-triple negative breast cancer and normal breast cells were analyzed by RT-PCR，immunocytochemistry and westernblot. DC were isolated and cultured from peripheral blood of healthy donors, the surface maker CD11c, CD80, CD86 and HLA-DR were analyzed with flow cytometry. Runx2 lentivirus were prepared to infect DC and divided in transfected group, negative group and blank group. Fluorescence microscope, PCR and Westernblot tested its transfection efficiency of all groups. DC and T cells were cultured together to induce CTL, ELISA was used to detect the secretion of IL-12 and IFN-γ. The killing ability of CTL were calculated by MTT.
Results: PCR results shown the ΔCT value of Runx2 in MDA –MB -231,MCF7 and MCF10A were 10.37±0.61, 11.86±0.59, 12.97±0.06（P < 0.05）respectively. Both immunocytochemistry and westernblot showed Runx2 were strongly positive in MDA –MB -231, but weaker in MCF7 and MCF10A.In this study, Runx2-lentivirus were prepared and transfected DC. Rich green flurescent confirmed that the Runx2 gene were successfully turned into mature DC .Then the transfected DC were co-cultured with T cells, and the secretion of IL-12、IFN-γ（pg/ml）were measured as 170.33±5.03、517.15±5.88. While the secretion of IL-12、IFN-γ were 72.30±3.05、171.57±1.37（P < 0.05）in non-transfected group. And the killing ratio of cytotoxic T cells of transfected group on triple negative breast cancer was 60.02±3.51 compared with 25.57±3.64（P < 0.05）of the control group.
Conclusions: Runx2 maybe a new target for triple negative breast cancer treatment. Runx2 can transduce DC successfully and the vaccine induce specific CTL and strong cytotoxicity against triple negative breast cancer.