BRD4对高表达PD-L1非小细胞肺癌细胞表达及增殖迁移的影响  

Effects of bromodomain-containing protein 4 on the expression, proliferation, and migration of non-small cell lung cancer with high expression of programmed death ligand 1

作　　者：姜剑松[1] 王晨路 朱曼旎 施敏骅[2] Jiang Jiansong;Wang Chenlu;Zhu Manni;Shi Minhua(Department of Pulmonary and Critical Care Medicine,Nantong First People′s Hospital,Nantong 226001,China;Department of Pulmonary and Critical Care Medicine,the Second Affiliated Hospital of Soochow University,Suzhou 215006,China)

机构地区：[1]南通市第一人民医院呼吸与危重症医学科,南通226001 [2]苏州大学第二附属医院呼吸与危重症医学科,苏州215006

出　　处：《国际呼吸杂志》2023年第5期569-575,共7页International Journal of Respiration

基　　金：南通市卫健委青年指令性课题项目(QN2022023)。

摘　　要：目的探讨BRD4对高表达程序性死亡受体配体1(PD-L1)非小细胞肺癌细胞PD-L1的表达及增殖迁移的影响。方法通过GEPIA中TCGA数据库, 获取有关于非小细胞肺癌患者的相关数据, 对高表达PD-L1、BRD4的非小细胞肺癌患者进行总生存率分析, 并对PD-L1和BRD4的表达情况进行相关性分析。PD-L1 mRNA及蛋白的表达水平, 选择高表达PD-L1的细胞株进行后续实验。分别用si-NC和si-BRD4转染A549细胞株, 获得NC组和敲低BRD4组。通过实时荧光聚合酶链反应和蛋白质印迹法分别检测2组的BRD4和PD-L1的mRNA及蛋白表达水平。集落形成实验检测细胞的增殖成瘤能力, 划痕实验检测细胞的迁移能力。分别加入DMSO和BRD4抑制剂JQ1获得对照组和JQ1组, 蛋白质印迹法和CCK-8实验检测2组0、12、24、36、48 h的A549细胞PD-L1蛋白的表达和细胞增殖水平。结果高表达BRD4、PD-L1的非小细胞肺癌患者中表现出低生存率(χ^(2)值分别为6.538、8.258, 均P<0.05)。PD-L1和BRD4的表达呈正相关(r=0.57, P<0.05, n=105)。A459细胞PD-L1 mRNA及蛋白的表达水平均高于H460、H1975细胞, 差异均有统计学意义(F值分别为2.58、2.67, 均P<0.05)。敲低BRD4组A549细胞中PD-L1的mRNA及蛋白表达水平低于NC组, 差异均有统计学意义(t值分别为6.37、7.54, 均P<0.05)。敲低BRD4的表达后, 非小细胞肺癌A549细胞的迁移能力、集落形成能力以及生长的能力较对照组降低。随着时间的增加, JQ1组PD-L1蛋白的表达水平逐步下调。自12 h开始, 2组PD-L1蛋白的表达水平比较差异均有统计学意义(t值分别为7.25、8.24、10.25、12.23, 均P<0.05)。CCK-8实验显示, 随着时间的增加, JQ1组A549细胞增殖水平逐步下调。自12 h开始, 2组A549细胞增殖水平差异均有统计学意义(t值分别为8.86、12.25、15.24、7.586, 均P<0.05)。结论抑制BRD4可以抑制高表达PD-L1非小细胞肺癌细胞PD-L1的表达及增殖迁移。Objective To explore the effect of bromodomain-containing protein 4(BRD4)on the expression,proliferation,and migration of non-small cell lung cancer(NSCLC)cells with high expression of programmed death ligand 1(PD-L1).Methods Relevant data on NSCLC patients were obtained through The Cancer Genome Atlas(TCGA)database in Gene Expression Profiling Interactive Analysis(GEPIA).The overall survival rate of NSCLC patients with high expression of PD-L1 and BRD4 was analyzed,and correlation analysis on the expression of BRD4 and PD-L1 was conducted.The mRNA and protein expression levels of PD-L1 were determined,and cell lines with high expression of PD-L1 were selected for subsequent experiments.A549 cells were transfected with si-NC and si-BRD4,respectively,to obtain NC group and BRD4 knock-down group.Subsequently,the mRNA and protein expression levels of BRD4 and PD-L1 in both groups were detected by real-time fluorescence polymerase chain reaction(RT-PCR)and Western blotting.Colony formation assay was used to detect the proliferation and tumorigenesis ability of cells,and scratch assay was used to detect the migration ability of cells.Meanwhile,the control group and JQ1 group were obtained by adding dimethyl sulfoxide(DMSO)and BRD4 inhibitor JQ1,respectively.Western blotting and Cell Counting Kit-8(CCK-8)assay were applied to detect the protein expression of PD-L1 and the proliferation of A549 cells of the two groups at 0,12,24,36,and 48 hours,respectively.Results Poor survival rate was observed in NSCLC patients with high expression of BRD4 or PD-L1(χ^(2) values were 6.538 and 8.258,respectively;allP<0.05).Positive correlation was observed between the expression of BRD4 and PD-L1(r=0.57,P<0.05,n=105).The mRNA and protein expression levels of PD-L1 in A459 cells were significantly higher than those in H460 and H1975 cells(Fvalues were 2.58 and 2.67,respectively;all P<0.05).The mRNA and protein expression levels of PD-L1 in A549 cells in BRD4 knock-down group were significantly lower than those in NC group(tvalues were

关 键 词：癌 非小细胞肺 程序性死亡受体配体1 BRD4 JQ1 

分 类 号：R734.2[医药卫生—肿瘤]