MiR-424-5p调控PD-1/PD-L1信号通路对弥漫大B细胞淋巴瘤细胞耐药性的影响  

Effect of MiR-424-5p on the Drug Resistance of Diffuse Large B-Cell Lymphoma Cells by Regulating PD-1/PD-L1 Signaling Pathway

作　　者：袁军[1] 韩虎 董巍[3] 王瑞仓[1] 郝洪岭[1] YUAN Jun;HAN Hu;DONG Wei;WANG Rui-Cang;HAO Hong-Ling(Department of Hematology,Hebei General Hospital,Shijiazhuang 050055,Hebei Province,China;Emergency Department,Hebei General Hospital,Shijiazhuang 050055,Hebei Province,China;Department of Cardiology,The Third Hospital of Shijiazhuang,Shijiazhuang 050000,Hebei Province,China)

机构地区：[1]河北省人民医院血液科,河北石家庄050055 [2]河北省人民医院急诊科,河北石家庄050055 [3]石家庄市第三医院心内科,河北石家庄050000

出　　处：《中国实验血液学杂志》2023年第1期96-103,共8页Journal of Experimental Hematology

基　　金：河北省医学科学研究所研究课题(20190282)。

摘　　要：目的:探讨微小RNA-424-5p(mi R-424-5p)调控程序性死亡受体-1(PD-1)/程序性死亡配体-1(PD-L1)信号通路对弥漫大B细胞淋巴瘤细胞耐药性的影响。方法:诱导人弥漫大B细胞淋巴瘤细胞系CRL2631细胞构建CRL2631-CHOP耐药细胞株。RT-q PCR和Western blot分别检测CRL2631细胞和CRL2631-CHOP细胞中mi R-424-5p、PD-L1 m RNA和蛋白及多元药物抗性基因-1(MDR-1)蛋白表达水平。双荧光素酶报告基因实验验证mi R-424-5p的靶标基因。使用mi RNA模拟/干扰技术和噻唑蓝法检测CRL2631细胞和CRL2631-CHOP细胞对CHOP方案化疗药物的耐药性。结果:与CRL2631细胞相比,CRL2631-CHOP细胞对CHOP方案化疗药物的耐药性显著增高,细胞中MDR-1蛋白水平(P<0.05)、PD-L1 m RNA和蛋白水平显著增高(均P<0.001),而mi R-424-5p相对水平显著降低(P<0.001)。双荧光素酶报告基因实验结果显示,PD-L1是mi R-424-5p下游直接靶标基因(P<0.001)。转染mi R-424-5p inhibitor后,CRL2631细胞对CHOP方案化疗药物的耐药性增高,且MDR-1蛋白水平(P<0.01)、PD-L1 m RNA和蛋白水平显著增高(均P<0.01);转染mi R-424-5p mimics后,CRL2631-CHOP细胞对CHOP药物的耐药性降低,且MDR-1蛋白水平(P<0.001)、PD-L1 m RNA和蛋白水平显著降低(均P<0.001);过表达PD-L1可逆转上调mi R-424-5p对PD-L1的抑制作用(P<0.001)。结论:下调mi R-424-5p通过调控PD-1/PD-L1信号通路增强了弥漫大B细胞淋巴瘤细胞的耐药性。Objective:To explore the effect of micro RNA-424-5p (mi R-424-5p) on the drug resistance of diffuse large B-cell lymphoma (DLBCL) cells by regulating the programmed death receptor-1 (PD-1)/programmed death ligand-1 (PD-L1) signaling pathway.Methods:Human DLBCL cell line CRL2631 cells were induced to construct CRL2631-CHOP resistant cell line.RT-q PCR and Western blot were used to detect the expression levels of mi R-424-5p,PD-L1 m RNA and protein,and multidrug resistance gene-1 (MDR-1) protein in CRL2631 cells and CRL2631-CHOP cells,respectively.The target genes of mi R-424-5p was verified by dual luciferase reporter assay.The mi RNA simulation/interference technology and thiazole blue (MTT) method were used to detect the resistance of CRL2631 cells and CRL2631-CHOP cells to CHOP.Results:Compared with CRL2631 cells,the drug resistance of CRL2631-CHOP cells to CHOP and the levels of MDR-1 protein (P<0.05),PD-L1 m RNA and protein in the cells were significantly increased (both P<0.001),while the relative level of mi R-424-5p was significantly reduced (P<0.001).The result of the dual luciferase reporter assay showed that PD-L1 was the direct downstream target gene of mi R-424-5p (P<0.001).After transfection of mi R-424-5p inhibitor,the resistance of CRL2631 cells to CHOP drugs increased,and the expression level of MDR-1 protein (P<0.01),PD-L1m RNA and protein also increased significantly (both P<0.01).After transfection of mi R-424-5p mimics,the resistance of CRL2631-CHOP cells to CHOP drugs decreased,and the expression level of MDR-1 protein (P<0.001),PD-L1 m RNA and protein also decreased significantly (both P<0.001).Overexpression of PD-L1 could reverse the inhibitory effect of upregulating mi R-424-5p on PD-L1 (P<0.001).Conclusion:Down-regulation of mi R-424-5p enhances the drug resistance of DLBCL cells by regulating the PD-1/PD-L1 signaling pathway.

关 键 词：微小RNA-424-5p 程序性死亡受体-1 程序性死亡配体-1 弥漫大B细胞淋巴瘤 耐药性 

分 类 号：R733.1[医药卫生—肿瘤]