NECTIN1缺失在食管鳞癌发生和发展中的作用及其机制  

Effect and Mechanism of NECTIN1 Deletion on the Occurrence and Progression of Esophageal Squamous Cell Carcinoma

作　　者：杨航 潘英杰 龙元凤 张佳怡 乔彦 刘云[1] 杨思芸 宋桂芹[1] 刘康 YANG Hang;PAN Yingjie;LONG Yuanfeng;ZHANG Jiayi;QIAO Yan;LIU Yun;YANG Siyun;SONG Guiqin;LIU Kang(Institute of Basic Medicine and Forensic Medicine,North Sichuan Medical College,Nanchong,637100;Nanchong Central Hospital,Beijing Anzhen Nanchong Hospital of Capital Medical University,Institute of Tissue Engineering and Stem Cells,Second Clinical Medical College of North Sichuan Medical College,Nanchong,637000)

机构地区：[1]川北医学院基础医学与法医学研究所,南充637100 [2]首都医科大学附属北京安贞医院南充医院南充市中心医院·川北医学院第二临床医学院组织工程与干细胞研究所,南充637000

出　　处：《基因组学与应用生物学》2025年第1期92-106,共15页Genomics and Applied Biology

基　　金：国家自然科学基金青年基金项目(82203851);四川省自然科学基金项目(2023NSFSC0731);南充市基础研究平台项目(23JCYJPT0028);南充市市校合作项目(22SXQT0334,20SXQT0325)共同资助。

摘　　要：本研究旨在探究黏连蛋白细胞黏附分子1(nectin cell adhesion molecule 1,NECTIN1)缺失对食管鳞癌(esophageal squamous cell carcinoma,ESCC)细胞发生和发展的影响及其分子作用机制。利用UCSC数据库分析食管癌(esophageal carcinoma,ESCA)中NECTIN1基因的线性拷贝数,通过Western blot和免疫组化染色分析NECTIN1在食管鳞癌细胞、食管鳞癌组织和癌旁组织中的蛋白水平。利用TISIDB数据库分析NECTIN1表达与食管癌临床特征的相关性。采用shRNA敲低NECTIN1在KYSE150和KYSE510细胞中的表达,利用CCK-8实验、克隆形成实验、细胞划痕和Transwell实验以及流式细胞术检测敲低NECTIN1对细胞功能的影响。对敲低NECTIN1的KYSE510细胞进行转录组测序,筛选差异表达基因;通过GO、KEGG富集分析,探讨NECTIN1可能影响的信号调控网络。结果表明,NECTIN1在部分食管癌患者中拷贝数减少,蛋白水平显著低于癌旁组织的(P<0.0001),NECTIN1低表达的食管鳞癌患者分化较差(P<0.0001),与临床晚期分期呈负相关。敲低NECTIN1基因后KYSE150和KYSE510细胞增殖、迁移和侵袭能力均增强(P<0.05),细胞S期增多,并使细胞凋亡减少(P<0.01)。在KYSE150细胞中敲低NECTIN1,进行转录组测序分析后发现115个基因显著上调、209个基因显著下调。KEGG分析表明,差异表达基因富集于肿瘤坏死因子(tumor necrosis factor,TNF)信号通路(P<0.01),通过GEPIA数据库分析发现,在ESCA患者中,该通路中的3个基因(IL1B、TNFAIP3、CSF2)的表达水平与NECTIN1呈显著正相关(P<0.05),与转录组测序结果相符。NECTIN1的缺失促进了ESCC细胞增殖、迁移和侵袭,其机制可能是通过TNF信号通路发挥抑癌作用,调控ESCC的发生与发展。This study aims to explore the impact of deletion of the nectin cell adhesion molecule 1(NECTIN1)on the occurrence and development of esophageal squamous cell carcinoma(ESCC)and its molecular mechanisms.The linear copy number of NECTIN1 gene in esophageal carcinoma(ESCA)was analyzed using the UCSC database,and the protein levels of NECTIN1 in ESCC and normal tissues were analyzed using Western blot and immunohistochemical staining.Use TISIDB database to analyze the correlation between NECTIN1 expression and clinical features of ESCA.Using shRNA to knock down the expression of NECTIN1 in KYSE150 and KYSE510 cells,the effects of knocking down NECTIN1 on cell function were detected using CCK-8 assay,clone formation assay,cell scratch and Transwell assay,and flow cytometry.Transcriptome sequencing was performed on KYSE510 cells with NECTIN1 knockdown to screen for differentially expressed genes.GO and KEGG enrichment analysis was used to explore the signal regulatory network that NECTIN1 may affect.The results indicated that the copy number of NECTIN1 was reduced in some esophageal cancer patients,and the protein level was significantly lower than that in adjacent tissues(P<0.0001).Patients with low expression of NECTIN1 in esophageal squamous cell carcinoma had poor differentiation(P<0.0001),which was negatively correlated with clinical advanced staging.Knocking down the NECTIN1 gene enhanced the proliferation,migration,and invasion abilities of KYSE150 and KYSE510 cells(P<0.05),increased the S phase of cells,and reduced cell apoptosis(P<0.01).Knocking down NECTIN1 in KYSE150 cells and conducting transcriptome sequencing analysis revealed significant upregulation of 115 genes and significant downregulation of 209 genes.KEGG analysis showed that differentially expressed genes were enriched in the tumor necrosis factor(TNF)signaling pathway(P<0.01).The expression levels of three genes IL1B,TNFAIP3 and CSF2 in this pathway were significantly positively correlated with NECTIN1 in ESCA patients(P<0.05),consistent with tr

关 键 词：黏连蛋白细胞黏附分子1(NECTIN1) 食管鳞癌(ESCC) 增殖 侵袭 肿瘤坏死因子(TNF)信号通路 

分 类 号：R735.1[医药卫生—肿瘤]