肾细胞癌PBRM1表达缺失抑制CD8^(+)T细胞浸润  

PBRM1 Deletion Inhibits CD 8^(+)T Cells Infiltration in Tumor Microenvironment of Renal Cell Carcinoma

作　　者：谢宏俊 阙滔滔 魏毅 范义增 侯涛 李磊[1] 谷孝云 徐珊[1] XIE Hong-jun;QUE Tao-tao;WEI Yi;FAN Yi-zeng;HOU Tao;LI Lei;GU Xiao-yun;XU Shan(The First Affiliated Hospital of Xi’an Jiaotong University,Key Laboratory of Environment and Genes Related to Diseases,Ministry of Education,Key Laboratory for Tumor Precision Medicine of Shaanxi Province,Xi’an Jiaotong University,Xi’an 710061,China;Shaanxi Health In-formation Center,Health Commission of Shaanxi Province,Xi’an 710002,China)

机构地区：[1]西安交通大学第一附属医院,环境和疾病相关基因教育部重点实验室肿瘤研究室,陕西省肿瘤精准医学重点实验室,陕西西安710061 [2]陕西省卫生健康信息中心,陕西西安710002

出　　处：《肿瘤学杂志》2022年第7期579-585,共7页Journal of Chinese Oncology

基　　金：国家自然科学基金(82072829);陕西省自然科学基础研究计划-面上项目(2021JM-265)。

摘　　要：[目的]探讨PBRM1调控CD8^(+)T在肾透明细胞癌组织中细胞浸润的分子机制。[方法]以肾透明细胞癌患者肿瘤临床标本为研究对象,探讨CD8^(+)T细胞在人肾癌中的浸润及其与临床病理特征的关系,以免疫印迹、荧光免疫、ELISA等分子生物学方法明确PBRM1调控CD8^(+)T细胞浸润的分子机制。免疫荧光检测CD63、EZH2及PHK26在细胞中的分布。免疫印迹检测IFN-γ,IL-6和TNF-α不同因子处理对肾癌细胞EZH2的诱导表达。实验结果经GraphPad Prism软件,t检验对两组间差异进行统计学分析。[结果]肾透明细胞癌组织免疫组化结果显示,在肿瘤组织中CD8^(+)T细胞浸润数量显著性下降(P<0.001),CD8^(+)T细胞浸润与患者总生存期呈正相关(P<0.001)。患者肿瘤组织PBRM1着色和CD8^(+)T细胞浸润数量结果显示:PBRM1表达与CD8^(+)T细胞浸润数量呈正相关(P<0.001)。IFN-γ,IL-6和TNF-α细胞因子促进EZH2在细胞表达升高,同时被外泌体包载被运输到受体细胞,外泌体EZH2进入受体细胞抑制CXCL9和CXCL10细胞因子分泌。[结论] PBRM1缺失激活exo-EZH2/CXCL9(10)信号轴抑制CD8^(+)T细胞肿瘤组织浸润。[Objective] To explore the molecular mechanism of PBRM1 in regulating CD8^(+)T cell infiltration in clear cell renal cell carcinoma tissue. [Methods] The expression of PBRM1 in renal clear cell carcinoma were detected by tissue microarray and its relation with CD8^(+)T cell infiltration in tumor microenvironment was analyzed. Western blot was also applied to detect EZH2 expression after IFN-γ, IL-6 and TNF-α treatment in renal cell carcinoma cells. Immunofluorescence was used to detect the distribution of CD63, EZH2 and PHK26 in cells. The effect of EZH2 on CXCL9/10 secretion was detected by ELISA assay. The Graph Pad Prism software was used for statistical analysis. [Results] Tissue microarray results showed that the infiltration density of CD8^(+)T cells in tumor tissues was lower than that in adjacent tissues(P<0.001). High CD8^(+)T cell infiltration was positively correlated with the overall survival of patients( P < 0. 001), and low PBRM1 protein expression was associated with decreased infiltration of CD8^(+)T cells(P <0.001).IFN-γ, IL-6 and TNF-α promoted the expression of EZH2 protein, which was packaged in exosomes and entered into recipient cells to inhibit CXCL9 and CXCL10 secretion. [Conclusion]PBRM1 deletion activates the exo-EZH2/CXCL9(10) signal axis and inhibits CD8^(+)T cell tumor tissue infiltration in renal cell carcinoma.

关 键 词：肾细胞癌 PBRM1 外泌体 CD8^(+)T细胞 EZH2 

分 类 号：R737.11[医药卫生—肿瘤]