CIK细胞对RNAi沉默PD-L1基因的肺腺癌A549细胞的体外抑制作用

目的:观察沉默肺癌A549细胞的程序性死亡受体1（programmed death ligand 1，PD-L1）基因表达后，CIK细胞对其体外抑癌的作用。方法:分离外周血单个核细胞(peripheral blood mononuclear cell，PBMC),加入细胞因子(cytokine，CK),体外细胞因子诱导为杀伤细胞(cytokine induced killer cells,CIK cells)。将已构建的PD-L1 shRNA 重组质粒 pGPU6/PD-L1，应用脂质体转染法转染入A549细胞。RT-PCR法检测PD-L1基因的表达；Western blotting法检测PD-L1蛋白的表达；CCK8法检测CIK细胞对各组A549细胞的杀伤率。结果:成功将pGPU6/PD-L1转染入A549细胞；RT-PCR结果显示pGPU6/PD-L1使A549细胞PD-L1基因表达水平降低；Western blotting 结果显示 pGPU6/PD-L1 转染A549细胞使 PD-L1蛋白表达减少；CCK8结果显示 pGPU6/PD-L1能够增强CIK细胞对A549细胞的体外抑瘤能力。结论:下调肺癌细胞A549的PD-L1 mRNA表达，MTT证明能够增强CIK细胞对其体外杀伤作用。

Antitumor effects of CIK cells on A549 cell lines silencing of PD-L1 by RNAi in vitro

Objective:Knockdown the programmed death ligand 1(PD-L1) in human lung adenocarcinoma cell line A549,to investigate antitumor effects of CIK on A549 cells in vitro.Methods:The peripheral blood mononuclear cell(PBMC) was isolated and added cytokine(CK).The cytokine induced killer cells(CIK cells) was induced.The PD-L1 shRNA plasmid pGPU6/PD-L1 has been constructed,transfected into A549 cells by liposome.The PD-L1 gene express was detected by Real-time Quantitative PCR,the PD-L1 protein expression of each cell group was detected by Western blot,CCK8 assay CIK cell killing rate for each group of A549 cells.Results:The A549 cells were successfully transfected by pGPU6/PD-L1.The result of real-time quantitative PCR showed that the PD-L1 gene was down-regulated by pGPU6/PD-L1.The significant down-regulation of PD-L1 expression caused by transfection of pGPU6/PD-L1 into A549 cells was detected by Western blotting.CCK8 assay showed pGPU6/PD-L1 could enhanced the antitumor effects of CIK cells to A549 cells in vitro.Conclusion:Reduced lung cancer cell line A549 PD-L1 expression,can enhance the antitumor effects of CIK cells to A549 cells in vitro.