miR-15a-5p靶向PD-L1对食管癌进展、预后与肿瘤免疫反应的影响  

MicroRNA-15a-5p targets programmed death-ligand 1 to impact progression,prognosis,and tumor immune response in esophageal cancer

作　　者：林亭呈 张婉君 Ting-Cheng Lin;Wan-Jun Zhang(Department of Pharmacy and Pharmacology,Taizhou Hospital of Zhejiang Province,Taizhou 318050,Zhejiang Province,China)

机构地区：[1]浙江省台州医院药剂科,浙江省台州市318050

出　　处：《世界华人消化杂志》2024年第9期678-685,共8页World Chinese Journal of Digestology

摘　　要：背景微核糖核酸(microRNA,miR)-15a-5p与细胞程序性死亡蛋白配体1(programmed death-ligand 1,PD-L1)在多种肿瘤进展、治疗与预后中发挥重要作用,但其在食管癌中的作用尚不完全清楚.目的研究miR-15a-5p与PD-L1表达对食管癌进展与预后的影响,并分析二者的靶向关系以及二者调控食管癌细胞生物学行为的机制.方法用癌症基因组图谱数据库、基因表达综合数据库和RNA相互作用组百科全书泛癌分析数据库分析食管癌中miR-15a-5p和PD-L1基因的表达,通过Kaplan-Meier生存分析研究miR-15a-5p和PD-L1的表达与食管癌无病生存期以及总生存期的关系.信息学和荧光活性实验检测miR-15a-5p与PD-L1的靶向关系.体外培养食管癌细胞,转染miR-15a-5p mimics或miR-NC后,分别用Western blot、流式细胞术、免疫荧光实验检测PD-L1表达.将转染miR-15a-5p mimics或miR-NC的食管癌细胞与CD8+T细胞共培养,流式细胞术检测干扰素-γ(interferon-γ,INF-γ)阳性CD8+T细胞比例,ELISA检测共培养体系中上清液中INF-γ和白细胞介素(interleukin,IL)-2含量.结果miR-15a-5p在食管癌组织中表达水平较癌旁非癌变组织降低(P<0.05).以miR-15a-5p表达水平的中位数为分界线,随T分期和肿瘤大小增加和淋巴结转移,miR-15a-5p低表达率增加(P<0.05);相比于miR-15a-5p低表达的食管癌患者,miR-15a-5p高表达显示出较长的无病生存期和总生存期(P<0.05),miR-15a-5p高表达同时PD-L1低表达的无病生存期和总生存期更长(P<0.05).在miR-15a-5p高表达的食管癌患者中PD-L1 mRNA多呈低表达.PD-L1为miR-15a-5p的靶基因.过表达miR-15a-5p抑制食管癌细胞中总PD-L1及膜PD-L1表达(P<0.05).在食管癌细胞中过表达miR-15a-5p可促进共培养体系中CD8+T细胞的激活(即:共培养体系中INF-γ阳性CD8+T细胞比例以及共培养体系中上清液中INF-γ和IL-2含量增加).结论miR-15a-5p可通过靶向PD-L1增强食管癌细胞的肿瘤免疫反应,且miR-15a-5p/PD-L1�BACKGROUND MicroRNA(miR)-15a-5p and programmed death-ligand 1(PD-L1)are pivotal in the progression,treatment,and prognosis of various cancer types;however,their involvement in esophageal cancer remains incompletely elucidated.AIM To investigate the effects of the expression of miR-15a-5p and PD-L1 on the progression and prognosis of esophageal cancer,to analyze the targeting relationship between the two,and to explore the mechanism underlying their regulatory effect on the biological behaviors of esophageal cancer cells.METHODS The expression of miR-15a-5p and PD-L1 genes in esophageal cancer was examined utilizing data from The Cancer Genome Atlas database,Gene Expression Omnibus database,and The Encyclopedia of RNA Interactomes pan-cancer analysis database.The relationship between the expression of miR-15a-5p and PD-L1 and disease-free interval and overall survival of esophageal cancer patients was studied by Kaplan-Meier survival analysis.The targeting relationship between miR-15a-5p and PD-L1 was detected by informatics and fluorescence activity experiments.After transfection of esophageal cancer cells with miR-15a-5p mimics or miR-NC,the expression of PD-L1 was detected by Western blot,flow cytometry,and immunofluorescence assay.The esophageal cancer cells transfected with miR-15a-5p mimics or miR-NC were co-cultured with CD8+T cells.Interferon-γ(INF-γ)-positive CD8+T cells were detected by flow cytometry,and INF-γand interleukin(IL)-2 contents in the supernatant of the co-culture system were detected by ELISA.RESULTS The expression level of miR-15a-5p in esophageal cancer tissue was lower than that in adjacent non-cancerous tissue(P<0.05).With the increase in T stage,tumor size,and occurrence of lymph node metastasis,there was a concomitant rise in the rate of low miR-15a-5p expression when using the median expression level of miR-15a-5p as the threshold(P<0.05).Compared with patients with esophageal cancer with low expression of miR-15a-5p,high expression of miR-15a-5p was associated with longer disease

关 键 词：miR-15a-5p 食管癌 PD-L1 预后 肿瘤免疫反应 

分 类 号：R735.1[医药卫生—肿瘤]