肝细胞癌中环氧合酶-2表达抑制常规1型树突状细胞的瘤内富集及免疫应答的研究
Cyclooxygenase-2 expressing hepatocellular carcinoma inhibits the infiltration and immune response of conventional type 1 dendritic cells
李煜子 1汪洋 1程倩 1李祖寅 1李照 1朱继业 1高杰1
作者信息
- 1. 北京大学人民医院肝胆外科,北京 100044
- 折叠
摘要
目的 探讨肝细胞癌(hepatocellular carcinoma,HCC)中环氧合酶-2(cyclooxygenase-2,COX-2)表达对常规1型树突状细胞(conventional type 1 dendritic cell,cDC1)的瘤内富集及免疫应答功能的影响.方法 采用免疫荧光染色检测2016年1月至2017年6月北京大学人民医院行肝切除术的111例HCC患者肿瘤组织中cDC1瘤内富集及COX-2表达水平,将HCC患者分为cDC1浸润型与cDC1缺失型,分析HCC细胞表达COX-2与cDC1瘤内浸润的相关性.利用HCC单细胞转录组测序数据检验COX-2编码基因PTGS2与cDC1亚群比例的相关性.回顾性分析HCC肿瘤组织中cDC1浸润情况与患者临床特征及预后的相关性.通过体外造血干细胞(HSC)向cDC1诱导分化及流式分选,获得HSC来源cDC1,与HCC细胞系共培养后进行转录组测序,并用COX-2选择性抑制剂塞来昔布抑制HCC细胞系的COX-2表达,通过FITC-葡聚糖摄取实验、流式检测、Luminex多因子检测探索HCC中COX-2表达对cDC1的抗原摄取、共刺激分子表达和细胞因子分泌功能的影响.结果 免疫荧光染色提示cDC1缺失型(n=73)的HCC肿瘤组织中COX-2表达水平显著高于cDC1浸润型(n=38)(P=0.004 2).单细胞转录组测序数据分析提示PTGS2高表达的HCC肿瘤组织中cDC1亚群比例显著降低.cDC1浸润型的HCC患者总体生存率(P=0.037)、无瘤生存率(P=0.048)优于cDC1缺失型.cDC1与HCC共培养后抗原摄取功能、共刺激分子表达、细胞因子分泌的免疫应答功能受到抑制,塞来昔布可改善cDC1的免疫应答功能.结论 HCC中cDC1瘤内富集与患者预后良好相关.HCC中表达COX-2可能抑制cDC1向瘤内浸润,抑制cDC1抗原摄取、共刺激分子表达及细胞因子分泌的免疫应答功能.靶向COX-2的抑制剂可逆转HCC对cDC1的功能抑制.
Abstract
Objective To investigate the influence of COX-2 expression in hepatocellular carcinoma(HCC)on the infiltration and immune response of conventional type 1 dendritic cells(cDC1).Methods Clinicopathological data from 111 HCC patients undergoing radical hepatectomy at Peking University People's Hospital from Jan 2016 to Jun 2017 were retrospectively analyzed.Immunofluorescence staining was employed to evaluate the cDC1 infiltration and COX-2 expression in tumor tissues.Patients were divided into two groups based on cDC1 infiltration:cDC1 enrichment and cDC1 depletion,and the correlation between COX-2 expression and cDC1 infiltration was analyzed.Single-cell sequencing of HCC tumor tissues was used to further investigate the correlation between PTGS2,the encoding gene of COX-2,and cDC1 infiltration.Hematopoietic stem cells(HSC)were utilized for in vitro generation of cDC1.HSC-derived cDC1s were sorted by FACS and cocultured with HCC cell line SNU423.Celecoxib,a selective COX-2 inhibitor,was used to suppress the COX-2 expression in HCC cell line SNU423.The functions of cDC1 were explored by FITC-dextran uptake assay,flow cytometry,and Luminex multiplex cytokine assay.Results COX-2 expression was significantly higher in the cDC1 depletion group(n=73)compared to the cDC1 enrichment group(n=38)(P=0.004 2).Patients with higher PTGS2 expression had significantly lower proportion of cDC1.Increased cDC1 infiltration in the HCC tumor microenvironment correlated with improved patient overall survival rates(P=0.037)and disease-free survival rates(P=0.048).Results from FITC-dextran uptake assay,flow cytometry,and Luminex assay indicated that cDC1 co-cultured with HCC showed significantly reduced antigen uptake function,co-stimulatory molecule expression,and cytokine secretion,but partially abrogated with celecoxib treatment.Conclusions The intratumoral infiltration of cDC1 is positively correlated with favorable prognosis in HCC patients.Elevated COX-2 expression in HCC impedes the intratumoral accumulation of cDC1 and compromises their immune response capabilities.COX-2 inhibitors hold promise for enhancing cDC1 function in HCC.
关键词
癌,肝细胞/树突细胞/环氧合酶-2/免疫应答Key words
Carcinoma,hepatocellular/Dendritic cells/COX-2/Immune response引用本文复制引用
基金项目
国家自然科学基金(81871291)
国家自然科学基金(82172693)
出版年
2024
NETL
NSTL
万方数据