HBEGF部分通过LIF上调胶质母细胞瘤细胞中PD-L1的表达水平  

HBEGF up-regulate expression level of PD-L1 in glioblastoma cells partly through LIF

作　　者：梁博[1] 王新军[1] 赵杨[2] 王建业[1] 周少龙[1] 杨卓[1] LIANG Bo;WANG Xin-jun;ZHAO Yang;WANG Jian-ye;ZHOU Shao-long;YANG Zhuo(Dept of Neurosurgery,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Dept of Pharmaceutics,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第五附属医院神经外科,河南郑州450052 [2]郑州大学第五附属医院药学部,河南郑州450052

出　　处：《中国药理学通报》2021年第11期1585-1592,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No81972361)。

摘　　要：目的探讨肝素结合表皮生长因子(heparin-binding epidermal growth factor,HBEGF)调控白血病抑制因子(leukemia inhibitory factor,LIF)上调胶质瘤细胞中程序性死亡因子配体1(programmed death ligand 1,PD-L1)的机制。方法Pearson分析原发性胶质瘤中HBEGF、LIF和PD-L1 mRNA相关性;U251、U87为研究对象,添加HBEGF中和抗体及敲除HBEGF后再添加HBEGF,RT-qPCR和蛋白免疫印迹检测细胞中LIF和PD-L1 mRNA和蛋白水平;HBEGF敲除细胞中添加HBEGF基础上添加LIF中和抗体检测PD-L1 mRNA和蛋白水平。结果Pearson法分析显示HBEGF、LIF、PD-L1 mRNA两两均呈正相关(P<0.05)。添加HBEGF中和抗体后,随着细胞传代,LIF、PD-L1 mRNA和蛋白水平逐渐降低(P<0.05)。敲除HBEGF中,LIF、PD-L1 mRNA和蛋白水平降低(P<0.05),添加HBEGF后,LIF、PD-L1 mRNA和蛋白水平升高(P<0.05)。敲除HBEGF中添加HBEGF后,PD-L1 mRNA和蛋白水平升高(P<0.05),再添加LIF中和抗体后,PD-L1 mRNA和蛋白水平降低(P<0.05)。结论胶质母细胞瘤中HBEGF部分诱导LIF是HBEGF维持PD-L1表达的一个中间过程。Aim To investigate the mechanism of heparin-binding epidermal growth factor(HBEGF)in up-regulating the expression of programmed death ligand 1(PD-L1)in glioma cells by regulating leukemia inhibitory factor(LIF).Methods Pearson analysis was used to analyze the correlation between HBEGF,LIF and PD-L1 mRNA in primary gliomas;U251 and U87 cells were treated with HBEGF neutralizing antibody and HBEGF was added after HBEGF knockout,the mRNA and protein levels of LIF and PD-L1 were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blot;HBEGF knockout cells were added with LIF neutralizing antibody on the basis of adding HBEGF,and the levels of PD-L1 mRNA and protein were detected.Results Pearson analysis showed that HBEGF,LIF,and PD-L1 mRNA were positively correlated(P<0.05).After adding HBEGF neutralizing antibody to U251 and U87 cells,the mRNA and protein levels of LIF and PD-L1 gradually decreased with cell passage(P<0.05).After HBEGF knockout,the mRNA and protein levels of LIF and PD-L1 decreased(P<0.05),and the mRNA and protein levels of LIF and PD-L1 increased after HBEGF was added(P<0.05).In HBEGF knockout cell lines,the levels of PD-L1 mRNA and protein were up-regulated after HBEGF was added(P<0.05),and the levels of PD-L1 mRNA and protein was down-regulated by LIF neutralizing antibody(P<0.05).Conclusion The partial induction of LIF by HBEGF in glioblastoma is an intermediate process for HBEGF to maintain PD-L1 expression.

关 键 词：胶质瘤细胞 肝素结合表皮生长因子 白血病抑制因子 程序性死亡因子配体1 敲除 作用机制 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学]