APE1通过NF-κB通路调节PD-L1在喉鳞状细胞癌中的表达  

APE1 Regulates the Expression of PD-L1 through the NF-κB Pathway in Laryngeal Squamous Cell Carcinoma

作　　者：杜国强[1] 王立银[1] 王晓凤[2] 李君[1] 张明明[3] 王瑾[1] DU Guoqiang;WANG Liyin;WANG Xiaofeng;LI Jun;ZHANG Mingming;WANG Jin(Otorhinolaryngological Department,Shengjing Hospital,China Medical University,Shenyang 110004,China;Emergency Department,Shengjing Hospital,China Medical University,Shenyang 110004,China;Pathology Department,Shengjing Hospital,China Medical University,Shenyang 110004,China)

机构地区：[1]中国医科大学附属盛京医院耳鼻咽喉科,沈阳110004 [2]中国医科大学附属盛京医院急诊科,沈阳110004 [3]中国医科大学附属盛京医院病理科,沈阳110004

出　　处：《中国医科大学学报》2018年第10期865-870,共6页Journal of China Medical University

基　　金：国家自然科学基金青年基金(81202126)

摘　　要：目的检测脱嘌呤/脱嘧啶核酸内切酶1 (APE1)和程序性死亡蛋白配体1 (PD-L1)在喉鳞状细胞癌中的表达,探讨两者的相关性。方法采用实时定量荧光PCR检测60例喉癌标本及30例癌旁黏膜组织中PD-L1、APE1基因的表达,并分析其与临床病理因素的关系;利用siRNA脂质体转染、脂多糖[LPS,核因子κB (NF-κB)信号通路激动剂]和PDTC (NF-κB信号通路抑制剂)干预Hep-2细胞系,采用Western blotting检测APE1、PD-L1、NF-κB、pNF-κB蛋白的表达。结果喉癌组织中APE1、PD-L1基因的表达显著高于癌旁组织(P <0.05),两者表达呈正相关(r=0.37,P <0.01)。PD-L1表达与喉癌发生部位有关,声门型喉癌组织中PD-L1mRNA表达水平显著高于声门上型和声门下型(P <0.05)。同时,喉癌组织中APE1 m RNA的表达与淋巴结转移与否具有相关性(P <0.05)。APE1-siRNA转染Hep-2细胞后,显著抑制细胞APE1、NF-κB、pNF-κB、PD-L1蛋白的表达水平(P <0.05)。在一定浓度范围内,LPS和PDTC干预Hep-2细胞后,PD-L1、NF-κB蛋白表达分别逐渐增高和逐渐降低(P <0.05)。而沉默Hep-2细胞中APE1基因表达后进行LPS干预,细胞NF-κB、PD-L1表达较未干预组和LPS干预组降低(P <0.05)。结论相对于声门下型和声门上型喉癌,声门型喉癌可能对PD1/PD-L1免疫检查点相关的免疫治疗相对敏感。APE1可能通过NF-κB信号通路调节PD-L1的表达。PD-L1和APE1表达的联合检测对抑制喉癌生长具有潜在的临床价值。Objective To investigate the expression and correlation of apurinic/apyrimidinic endonuclease 1(APE1)and programmed death ligand 1(PD-L1)in human laryngeal squamous cell carcinoma(LSCC).Methods We collected 60 LSCC tissues and 30 adjacent mucosal tissues and detected expressions of PD-L1 and APE1 and analyzed their correlation with each other and with clinicopathological factors.Hep-2 cells were transfected with siRNA liposomes,lipopolysaccharides[nuclear factorκB(NF-κB)agonists],and PDTC(NF-κB inhibitor).The expressions of APE1,PD-L1,NF-κB,and pNF-κB were detected by Western blotting.Results The expressions of APE1 and PD-L1 mRNA were significantly higher in LSCC tissues than in adjacent tissues(P<0.05).There was a positive correlation between APE1 and PD-L1 mRNA expressions in LSCC tissues(r=0.37,P<0.01).The expression of PD-L1 was related to the tumor location,and the level of PD-L1 mRNA was significantly higher in glottic carcinoma than in subglottic and supraglottic carcinomas(P<0.05).APE1 mRNA expression was associated with lymphatic metastasis(P<0.05).The levels of APE1,NF-κB,pNF-κB,and PD-L1 were down-regulated in Hep-2 cells transfected by APE1-siRNA(P<0.05).PD-L1 and NF-κB were increased and decreased after treatment with lipopolysaccharides and PDTC in a certain concentration range,respectively.In APE1-siRNA transfected cells treated with lipopolysaccharides,NF-κB and PD-L1 were down-regulated compared with the control group and the lipopolysaccharide-only group(P<0.05).Conclusion Compared with subglottic and supraglottic carcinomas,glottic carcinoma may show a better response to PD-1/PD-L1-based immunotherapy.APE1 regulates PD-L1 expression through the NF-κB signaling pathway.Combined determination of PD-L1 and APE1 expressions may have potential clinical significance in inhibiting LSCC growth.

关 键 词：喉鳞状细胞癌 脱嘌呤/脱嘧啶核酸内切酶1 程序性死亡蛋白配体1 核因子ΚB信号通路 HEP-2细胞 

分 类 号：R767.1[医药卫生—耳鼻咽喉科]