Human dendritic cells transfected with MUC1 mRNA induce lethal effect of cytotoxic T-lymphocytes on non-small cell lung cancer in vitro / 中国病理生理杂志

ABSTRACT

AIM:To investigate the specific anti-tumor effects of mature dendritic cells ( DCs) transfected with amplified mucin 1 ( MUC1) mRNA in vitro.METHODS:DCs separated and purified from the peripheral blood mononu-clear cells were induced in vitro and then identified by flow cytometry .pcDNA3.1(+)-MUC1 plasmid was constructed and was able to transcribe MUC1 mRNA in vitro.The MUC1 mRNA was transfected into DCs by electroporation .MUC1-trans-fected DCs were used to induce T cells to be cytotoxic T-lymphocytes .Quantitative real-time PCR was performed to assess MUC1 mRNA expression in transfected DCs .The proliferation of T cells was examined by MTT assay .The proportion of CD8 +cells in the T cells was determined by flow cytometry and the specific cytotoxicity was measured by LDH assay .The secretion of IFN-γwas detected by ELISA .RESULTS: The marker gene expression in the DCs transfected with MUC 1 mRNA was significantly increased compared with control group , peaking at 24 h.The transfection group showed the higher capacity to stimulate the proliferation of T cells compared with control group when the ratio of DCs to T cells was 1∶10.The proportion of CD8 +cells in transfection group was higher than that in control group .The lethal effect of special cytotoxic T-lymphocytes on target cells in transfection group was stronger than that in control group .The level of IFN-γin the cell su-pernatant of transfection group was higher than that in control group .CONCLUSION:DCs plus MUC1 mRNA by electri-cal transfection induces specific anti-tumor effects , which provides an experiment evidence of using MUC 1 as a target for immunotherapeutic strategy against non-small cell lung cancer .