ZBED2通过糖酵解代谢诱导肝细胞癌中PD-L1表达促进免疫逃逸的机制研究  

Mechanism of immune escape induced by ZBED 2 induced PD-L 1 expression in hepatocellular carcinoma through glycolysis metabolism

作　　者：黄金石[1] 丁亚亭[1] 曹建中[1] HUANG Jinshi;DING Yating;CAO Jianzhong(Pharmacy Department of Affiliated Hospital of Nantong University,Nantong 226000,China)

机构地区：[1]南通大学附属医院药学部,南通226000

出　　处：《中国免疫学杂志》2025年第2期367-373,共7页Chinese Journal of Immunology

摘　　要：目的:探讨锌指BED结构域含蛋白2(ZBED2)通过糖酵解通路对肝细胞癌(HCC)免疫逃逸的影响及潜在机制。方法:生物信息学数据库分析ZBED2在HCC组织中的表达情况及二者的结合位点,分析ZBED2调控的通路,以及ZBED2与糖酵解基因的相关性。qPCR和Western blot检测ZBED2及程序性细胞死亡配体1(PD-L1)在HCC细胞中的表达,MTT检测细胞活力,细胞毒性实验检测CD8+T细胞毒性,ELISA检测细胞因子表达;细胞外流量分析仪检测胞外酸化率(ECAR)和耗氧率(OCR),qPCR检测糖酵解相关基因表达,试剂盒检测糖酵解指标,免疫组织化学染色检测肿瘤组织中CD8^(+)T细胞表达。结果:ZBED2在HCC中表达上调,过表达ZBED2可促进PD-L1表达,抑制CD8^(+)T细胞对HCC细胞的毒性。过表达ZBED2通过激活糖酵解通路抑制HCC中CD8^(+)T细胞活性,进一步添加糖酵解抑制剂2-脱氧-D-葡萄糖(2-DG)减弱了以上结果。体内实验发现,敲低ZBED2可抑制小鼠肿瘤生长及PD-L1表达,促进体内CD8+T细胞浸润。结论:ZBED2通过糖酵解代谢诱导HCC中PDL1表达,促进免疫逃逸。Objective:To investigate the effect of zine finger BED domain-containing protein 2(ZBED2)on immune escape of hepatocellular carcinoma(HCC)through glycolysis pathway and its potential mechanism.Methods:Expression of ZBED2 in HCC tissues and binding sites of them were analyzed in bioinformatics database,the pathway regulated by ZBED2 was analyzed,as well as the correlation between ZBED2 and glycolysis genes.qPCR and Western blot were used to detected expressions of ZBED2 and programmed death-ligand 1(PD-L1)in HCC cells,cell viability was detected by MTT,toxicity of CD8+T cells was detected by cytotoxicity assay,and cytokine expression was detected by ELISA.Extracellular acidification rate(ECAR)and oxygen consumption rate(OCR)were detected by extracellular flow analyzer,glycolytic gene expression was detected by qPCR,and glycolytic index was detected by kit.Expression of CD8^(+)T cell in tumor tissues was detected by immunohistochemical staining.Results:ZBED2 was up-regulated in HCC,overexpression of ZBED2 could promote expression of PD-L1,while inhibit cytotoxicity of CD8^(+)T cells to HCC.Overexpression of ZBED2 inhibited CD8^(+)T cell activity in HCC by activating glycolysis pathway,and further addition of glycolysis inhibitor 2-DG attenuated the above results.In vivo experiments showed that ZBED2 knockdown inhibited tumor growth,inhibited PD-L1 expression,while promoted CD8+T cell infiltration in vivo.Conclusion:ZBED2 induces expression of PD-L1 in HCC through glycolytic metabolism,and promotes immune escape.

关 键 词：锌指BED结构域含蛋白2 糖酵解 肝细胞癌 免疫逃逸 

分 类 号：R392.12[医药卫生—免疫学]