Wang Zhen, Huang Wen, Cen Bohong, Wei Yuanyi, Liao Lumin, Li Guoxian, Ji Aimin
Department of Pharmacy, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, China.
R&D Center, Nanjing Pharmaceutical Factory Co., Ltd., Nanjing 210007, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2018 Jul 30;38(7):800-806. doi: 10.3969/j.issn.1673-4254.2018.07.05.
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.
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.
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).
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.
研究小干扰RNA（siRNA）介导的人胶质瘤细胞程序性细胞死亡配体1（PD-L1）沉默对人CD8+T淋巴细胞对修饰肿瘤细胞细胞毒性的影响。
设计靶向PD-L1基因的siRNA序列，通过脂质体2000转染到人胶质瘤U87 MG细胞中，采用RT-qPCR、蛋白质印迹法和流式细胞术验证基因沉默效果。将转染后的细胞与人CD8+T淋巴细胞共培养，用流式细胞术分析肿瘤细胞的凋亡情况。
该siRNA序列在U87 MG细胞的mRNA和蛋白质水平均显示出较强的PD-L1基因沉默效果。与对照细胞相比，转染后的U87 MG细胞对人CD8+T细胞的细胞毒性敏感性显著增加，共培养时增殖活性明显降低（P<0.05）。
用靶向PD-L1的特异性siRNA转染人胶质瘤U87 MG细胞可明显增强共培养时人T淋巴细胞的毒性。
