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.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb;31(1):96-103. doi: 10.19746/j.cnki.issn.1009-2137.2023.01.015.
To explore the effect of microRNA-424-5p (miR-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.
Human DLBCL cell line CRL2631 cells were induced to construct CRL2631-CHOP resistant cell line. RT-qPCR and Western blot were used to detect the expression levels of , PD-L1 mRNA and protein, and multidrug resistance gene-1 (MDR-1) protein in CRL2631 cells and CRL2631-CHOP cells, respectively. The target genes of was verified by dual luciferase reporter assay. The miRNA simulation/interference technology and thiazole blue (MTT) method were used to detect the resistance of CRL2631 cells and CRL2631-CHOP cells to CHOP.
Compared with CRL2631 cells, the drug resistance of CRL2631-CHOP cells to CHOP and the levels of MDR-1 protein (<0.05), PD-L1 mRNA and protein in the cells were significantly increased (both <0.001), while the relative level of was significantly reduced (<0.001). The result of the dual luciferase reporter assay showed that PD-L1 was the direct downstream target gene of (<0.001). After transfection of inhibitor, the resistance of CRL2631 cells to CHOP drugs increased, and the expression level of MDR-1 protein (<0.01), PD-L1 mRNA and protein also increased significantly (both <0.01). After transfection of mimics, the resistance of CRL2631-CHOP cells to CHOP drugs decreased, and the expression level of MDR-1 protein (<0.001), PD-L1 mRNA and protein also decreased significantly (both <0.001). Overexpression of PD-L1 could reverse the inhibitory effect of upregulating on PD-L1 (<0.001).
Down-regulation of enhances the drug resistance of DLBCL cells by regulating the PD-1/PD-L1 signaling pathway.
通过调节程序性死亡受体-1（PD-1）/程序性死亡配体-1（PD-L1）信号通路，探讨微小RNA-424-5p（miR-424-5p）对弥漫性大B细胞淋巴瘤（DLBCL）细胞耐药性的影响。
诱导人DLBCL细胞系CRL2631细胞构建CRL2631-CHOP耐药细胞系。分别采用RT-qPCR和蛋白质印迹法检测CRL2631细胞和CRL2631-CHOP细胞中miR-424-5p、PD-L1 mRNA和蛋白以及多药耐药基因-1（MDR-1）蛋白的表达水平。通过双荧光素酶报告基因检测法验证miR-424-5p的靶基因。采用miRNA模拟/干扰技术和噻唑蓝（MTT）法检测CRL2631细胞和CRL2631-CHOP细胞对CHOP的耐药性。
与CRL2631细胞相比，CRL2631-CHOP细胞对CHOP的耐药性以及细胞中MDR-1蛋白水平（<0.05）、PD-L1 mRNA和蛋白水平均显著升高（均<0.001），而miR-424-5p的相对水平显著降低（<0.001）。双荧光素酶报告基因检测结果显示，PD-L1是miR-424-5p的直接下游靶基因（<0.001）。转染miR-424-5p抑制剂后，CRL2631细胞对CHOP药物的耐药性增加，MDR-1蛋白水平（<0.01）、PD-L1 mRNA和蛋白水平也显著升高（均<0.01）。转染miR-424-5p模拟物后，CRL2631-CHOP细胞对CHOP药物的耐药性降低，MDR-1蛋白水平（<0.001）、PD-L1 mRNA和蛋白水平也显著降低（均<0.001）。过表达PD-L1可逆转上调miR-424-5p对PD-L1的抑制作用（<0.001）。
下调miR-424-5p通过调节PD-1/PD-L1信号通路增强DLBCL细胞的耐药性。
