芦可替尼对人红白血病细胞免疫检查点分子表达的影响  

Effect of ruxolitinib on immune checkpoint molecule expression in human erythroleukemia cells

作　　者：付建珠 成志勇[1] 张朝 张丽军[2] 刘培 耿娜[1] FU Jianzhu;CHENG Zhiyong;ZHANG Zhao;ZHANG Lijun;LIU Pei;GENG Na(Department of Hematology,No.1 Hospital of Baoding,Baoding Hebei 071000;Postgraduate Research Institute,Chengde Medical College,Chengde Hebei 067000,China)

机构地区：[1]保定市第一医院血液内科,河北保定071000 [2]承德医学院研究生院,河北承德067000

出　　处：《临床与病理杂志》2024年第7期927-934,共8页Journal of Clinical and Pathological Research

基　　金：河北省重点研发计划(223777105D)。

摘　　要：目的:芦可替尼通过抑制酪氨酸蛋白激酶2(tyrosine protein kinase 2,JAK2)通路来抑制人红白血病(human erythroleukemia,HEL)细胞的血管新生及迁移能力,而HEL细胞中程序性死亡受体1(programmed death-1,PD-1)、程序性死亡受体配体1(programmed death-ligand 1,PD-L1)的表达及芦可替尼对其表达的影响尚无相关报道。本研究探讨芦可替尼对JAK2 V617F突变阳性HEL细胞增殖、凋亡以及免疫检查点分子PD-1、PD-L1表达的影响。方法:用不同浓度的芦可替尼处理HEL细胞,细胞增殖检测试剂盒(cell counting kit-8,CCK-8)检测细胞活力,流式细胞术检测HEL细胞凋亡率,荧光定量PCR检测JAK2、PD-1及PD-L1 mRNA表达水平,蛋白质印迹法检测p-JAK2、PD-1、PD-L1蛋白质表达水平。流式细胞术分析淋巴细胞与HEL细胞共培养48 h后HEL细胞PD-1、PD-L1的表达和淋巴细胞PD-1的表达及调节性T细胞比例。结果:不同浓度芦可替尼能够抑制HEL细胞增殖,并与时间和剂量呈正相关。芦可替尼能够降低JAK2、PD-1、PD-L1 mRNA及蛋白质表达水平,并与时间和剂量呈正相关。结论:芦可替尼可能通过特异性抑制JAK2信号通路降低HEL细胞PD-1、PD-L1的表达。Objective:Ruxolitinib inhibits angiogenesis and migration in human erythroleukemia(HEL)cells by suppressing the tyrosine protein kinase 2(JAK2)pathway.However,the expression of programmed death-1(PD-1)and programmed death-ligand 1(PD-L1)in HEL cells,as well as the effect of ruxolitinib on their expression,has not been reported.This study aims to investigate the impact of ruxolitinib on the proliferation,apoptosis,and expression of immune checkpoint molecules PD-1 and PD-L1 in JAK2V617F-mutated human erythroleukemia HEL cells.Methods:HEL cells were treated with different concentrations of ruxolitinib.Cell viability was measured using the cell counting kit-8(CCK-8)assay,and flow cytometry was employed to detect the apoptosis rate of HEL cells.RT-PCR was used to detect the mRNA expression levels of JAK2,PD-1,and PD-L1.Western blotting was used to detect the expression of phosphorylated JAK2(p-JAK2),PD-1,and PD-L1 proteins.Additionally,flow cytometry was used to evaluate the expressions of PD-1 and PD-L1 after 48 hours of co-culturing of lymphocytes and HEL cells.Results:Ruxolitinib inhibited the proliferation of HEL cells in a time-and dose-dependent manner.It also reduced the mRNA and protein expression levels of JAK2,PD-1,and PD-L1,with these effects being positively correlated with time and dose.Conclusion:Ruxolitinib may reduce PD-1 and PD-L1 expression in HEL cells by specifically inhibiting the JAK2 signaling pathway.

关 键 词：程序性死亡受体1 程序性死亡受体配体1 人红白血病细胞 芦可替尼 

分 类 号：R733.7[医药卫生—肿瘤]