[Small interfering RNA-mediated programmed cell death-ligand 1 silencing in human glioma cells enhances human CD8 T lymphocyte cytotoxicity ].

OBJECTIVE

To investigate the effect of small interfering RNA (siRNA)-mediated silencing of programmed cell deathligand 1 (PD-L1) in human glioma cells on the cytotoxicity of human CD8T lymphocytes against the modified tumor cells.

METHODS

A siRNA sequence targeting PD-L1 gene was designed and transfected into human glioma U87 MG cells via lipofectamine 2000, and the gene silencing effect was validated using RT-qPCR, Western blotting, and flow cytometry. The transfected cells were co-cultured with human CD8T lymphocytes, and the apoptosis of the tumor cells was analyzed with flow cytometry.

RESULTS

The siRNA sequence showed strong PD-L1 gene-silencing effect at both mRNA and protein levels in U87 MG cells. Compared with the control cells, the transfected U87 MG cells showed significantly increased vulnerability to the cytotoxicity of human CD8T cells and an obvious reduction of proliferative activity in the co-culture ( < 0.05).

CONCLUSIONS

Transfection of human glioma U87 MG cells with the specific siRNA targeting PD-L1 obviously enhances the toxicity of human T lymphocytes in the co-culture.