程序性死亡配体-1抑制剂对神经胶质瘤细胞的抑制作用及其机制  

Inhibitory effect of programmed cell death ligand-1 inhibitor on neuroglioma cells and its mechanism

作　　者：申法政[1] 崔士娟 梁甲宁 王向阳 孟磊[1] 马继伟[1] 赵新利[1] Shen Fazheng;Cui Shjuan;Liang Jianing;Wang Xiangyang;Meng Lei;Ma Jiwei;Zhao Xinli(Department of Neurosurgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100 China;Department of Tuberculosis Internal Medicine,the First Affiliated Hospital of Xinxiang Medical Medical University,Weihui 453100,Chino)

机构地区：[1]新乡医学院第一附属医院神经外科,卫辉453100 [2]新乡医学院第一附属医院结核内科,卫辉453100

出　　处：《中华实验外科杂志》2024年第7期1498-1501,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划项目(LHGJ20210515)。

摘　　要：目的探讨程序性死亡配体-1(PD-L1)抑制剂对神经胶质瘤细胞恶性生物学行为的抑制作用及其作用机制。方法选取人正常星形胶质细胞HA1800和人神经胶质瘤细胞U87-MG、U251-MG作为研究对象,实时荧光定量聚合酶链反应检测HA1800、U87-MG、U251-MG细胞PD-L1 mRNA表达水平;U87-MG细胞分为两组,分别是添加PD-L1抑制剂的U87-MG细胞(实验组)和未经PD-L1抑制剂处理的U87-MG细胞(对照组)。采用细胞计数试剂盒(CCK-8)检测实验组和对照组细胞的增殖活力;采用Transwell小室检测实验组和对照组细胞的迁移与侵袭能力;采用蛋白免疫印迹检测实验组和对照组细胞PD-L1、信号传导及转录激活蛋白1(STAT1)和信号传导及转录激活蛋白3(STAT3)磷酸化水平。组间比较采用t检验。结果U87-MG细胞(1.73±0.08)和U251-MG细胞(1.62±0.06)中PD-L1mRNA的表达水平明显高于HA1800细胞(0.49±0.05),差异有统计学意义(t=31.91、37.70,P<0.05)。实验组细胞(0.52±0.05.0.82±0.04)在第48、72小时的吸光度(A)值明显低于对照组细胞(0.80±0.05、1.32±0.06),差异有统计学意义(t=9.49、17.94,P<0.05)。实验组细胞迁移细胞数[(73.33±5.35)个]和侵袭细胞数[(46.17±5.12)个]明显低于对照细胞[(135.17±7.52)、(88.50±6.77)个],差异有统计学意义(t=16.41、12.21,P<0.05)。实验组细胞PD-L1蛋白水平(0.88±0.05)、STAT3磷酸化水平(0.44±0.05)明显低于对照组细胞(1.88±0.06、1.24±0.05)差异有统计学意义(t=30.34、26.73,P<0.05)。实验组细胞STAT1磷酸化水平(1.32±0.08)明显高于对照组细胞(0.51±0.05),差异有统计学意义(t=20.43,P<0.05)。结论PD-L1抑制剂可通过上调磷酸化STAT1水平、下调STAT3磷酸化水平,抑制神经胶质瘤细胞的增殖、迁移和侵袭。Objective eTo investigate the inhibitory effect of programmed cell death ligand-1(PD-L1)inhibitor on malignant biological behavior of neuroglioma cells and its mechanism.Methods Human normal astrocytes HA1800 and human neuroglioma cells(U87-MG,U251-MG)were selected for the study.Real-time fluorescence quantitative polymerase chain reaction was used to detect the PD-Ll mRNA expression in HA1800,U87-MG,and U251-MG cells.U87-MG cells were divided into two groups:U87-MG cells with the addition of PD-L1 inhibitor(experimental group)and U87-MG cells without PD-L1 inhibitor treatment(control group).The cell proliferation viability in the experimental and control groups was detected using a cell counting kit-8(CCK-8)assay;Transwell was used to detect the migration and in-vasion ability of the experimental group and the control group.The protein expression of PD-Ll,phospho-rylation signal transducer and activators of transcription 1(STAT1),and phosphorylation signal transducer and activators of transcription 3(STAT3)in the experimental group and control group was detected by Western blotting.The t-test was used for comparison between groups.Results The expression of PD-Li mRNA in U87-MG cells(1.73±0.08)and U251-MG cells(1.62±0.06)was significantly higher than that in HA1800 cells(0.49±0.05,t=31.91,37.70,P<0.05).The absorbance(A)value of the experi-mental group(0.52±0.05,0.82±0.04)at 48 h and 72 h was significantly lower than that of the control group(0.80±0.05,1.32±0.06,t=9.49,17.94,P<0.05).The number of migrating cells[(73.33±5.35)]and invasive cells[(46.17±5.12)]in the experimental group was significantly less than that in the control group[(135.17±7.52)and(88.50±6.77),t=16.41,12.21,P<0.05].The protein expression of PD-L1(0.88±0.05)and phosphorylated STAT3(0.44±0.05)in experimental group was significantly lower than that in control group(1.88±0.06,1.24±0.05,t=30.34,26.73,P<0.05).The phosphorylated STAT1 in experimental group(1.32±0.08)was significantly higher than that in control group(0.51±0.05,t=20.43

关 键 词：程序性死亡配体-1抑制剂 神经胶质瘤 增殖 迁移 侵袭 

分 类 号：R739.4[医药卫生—肿瘤]