Effect of Programmed Death Ligand-1 signal on activity and apoptosis of Jurkat cell.
Shuang Li, Xu Lan, Xiuchun Zhang, Zhu Yun-lan, Yuedan Wang
Abstract
Shuang Li, Xu Lan, Xiuchun Zhang, Zhu Yun-lan, Yuedan Wang
Abstract
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.
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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.
Key concepts: Jurkat cells, Apoptosis, Small interfering RNA, Cell, Flow cytometry, Molecular biology, Cell biology, Biology