中文摘要：

目的：探讨PD-L1（Programed Death Ligand-1）表达的调控以及其对Jurkat细胞增殖分化及凋亡的影响。方法：通过IFN-γ在体外诱导Jurkat细胞使PD-L1表达上调,并设计针对PD-L1的siRNA对Jurkat细胞进行转染抑制PD-L1的表达,通过RT-PCR、PCR测定Jurkat细胞分泌IL-2的变化;流式细胞术测定Jurkat细胞的凋亡情况。结果：运用β-actin进行半定量后,根据相应的模板量进行PCR,在设计的3条针对PD-L1的siRNA（siRNA-A,siRNA-B,siRNA-C）中选出一条可以高效抑制PD-L1表达的即siRNA-A,siRNA-A终浓度80nM转染Jurkat细胞24小时后PD-L1的表达明显抑制。使用IFN-γ终浓度2000U/ml作用于Jurkat细胞24小时后PD-L1表达明显增高,通过RT-PCR、PCR观察到PD-L1表达高低与Jurkat细胞分泌IL-2水平呈负相关,流式细胞术显示PD-L1表达增强能降低Jurkat细胞凋亡。结论：体外合成的针对PD-L1基因特异性siRNA,转染Jurkat细胞后可特异性抑制PD-L1的表达。IFN-γ在体外可刺激Jurkat细胞PD-L1表达增高。PD-L1信号在体外对Jurkat细胞分泌IL-2呈负相关,并且PD-L1能抑制Jurkat细胞凋亡。

英文摘要：

Objective：To investigate the expression regulation of PD-L1（Programed Death Ligand-1） in Jurkat cell and correlation between the expression of PD-L1（Programed Death Ligand-1,B7-H1） and the activity and apoptosis of Jurkat cell.Methods：IFN-γ and the siRNA（small interfering RNA） targeting to PD-L1 were used to control the expression of PD-L1 in the Jurkat cell.RT-PCR,PCR were used to measure the expression of IL-2 and flow cytometry was used to detected the apoptosis of Jurkat cell.Results：Semi-quantitative β-actin,according template amount to PCR,was used to choose siRNA-A from three different siRNA（siRNA-A,siRNA-B,siRNA-C） which can efficiently and specifically inhibit the expression of PD-L1 in Jurkat Cell.80nM siRNA-A transfect Jurkat Cell,after 24h,the expression of PD-L1 decrease obviously.2000U/ml IFN-γ induced Jurkat Cell,24h later,PD-L1 up-regulated.The expression of PD-L1 in Jurkat Cell negatively correlates with the level of IL-2 secreted by Jurkat Cell by RT-PCR and PCR.And the result from flow cytometry showed increased expression of PD-L1 can reduce the apoptosis of Jurkat Cell.Conclusions：Synthesized siRNA for PD-L1,after transfecting Jurkat cell,could specifically inhibit the expression of PD-L1.IFN-γ in vitro could stimulate Jurkat cell and increase the expression of PD-L1.PD-L1 signal negatively correlates with IL-2 secreted by Jurkat cell,and PD-L1 could inhibit the apoptosis of Jurkat cell.