转录因子En1通过调控Hedgehog信号通路促进食管鳞状细胞癌细胞增殖和迁移  

En1 promotes cell proliferation and migration via Hedgehog signaling pathway in esophageal squamous cell carcinoma

作　　者：赵宁 巩彤阳 魏子超 从计 刘芝华 陈洪岩 Zhao Ning;Gong Tongyang;Wei Zichao;Cong Ji;Liu Zhihua;Chen Hongyan(Key Laboratory of Molecular Oncology,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academic of Medical Sciences and Peking Union Medical College,Beijing 100021,China)

机构地区：[1]国家癌症中心、国家肿瘤临床医学研究中心、中国医学科学院北京协和医学院肿瘤医院分子肿瘤学国家重点实验室,北京100021

出　　处：《中华肿瘤杂志》2024年第2期99-107,共9页Chinese Journal of Oncology

摘　　要：目的探讨转录因子En1在食管鳞状细胞癌(ESCC)细胞中的功能及机制。方法利用癌症基因组图谱数据库(TCGA)中9 397例泛癌患者的En1表达和总生存资料、4 349例泛癌患者的En1表达和无进展生存资料, 分析泛癌中En1表达水平与患者预后的关系。利用基因表达综合数据库(GEO)的53对和国家基因组科学数据中心-组学原始数据归档库(NGDC-GSA)的155对ESCC组织和配对癌旁组织的基因表达资料分析ESCC组织中En1的表达水平。以慢病毒系统介导ESCC细胞KYSE180和KYSE450中En1基因敲降, 采用细胞计数试剂盒8法和克隆形成实验检测细胞的增殖能力, Transwell实验检测细胞的迁移能力, 采用裸鼠皮下移植瘤实验检测En1对ESCC细胞体内肿瘤生长的影响。采用实时荧光定量聚合酶链反应(RT-qPCR)检测细胞中En1及Hedgehog通路主要调控因子胶质瘤相关癌基因家族锌指1(GLI1)、GLI2和平滑蛋白(SMO)的表达。结果来自TCGA数据库的泛癌样本资料显示, En1低表达患者的总生存时间和无进展生存时间均比En1高表达患者更长(均P<0.001)。来自GEO和NGDC-GSA数据库的资料显示, ESCC组织中En1的表达水平高于配对癌旁组织(均P<0.001)。功能研究显示, 与shNC组相比, 敲降En1能显著抑制KYSE180和KYSE450细胞的增殖(均P<0.001)、抑制克隆形成[KYSE180细胞:shEn1#1组和shEn1#2组的克隆形成数分别为(138.33±23.07)个和(127.00±19.70)个, 均低于shNC组的(340.67±12.06)个(均P<0.001);KYSE450细胞:shEn1#1组和shEn1#2组的克隆形成数分别为(65.33±2.52)个和(9.00±3.00)个, 均低于shNC组的(139.00±13.00)个(均P<0.001)]、抑制迁移[KYSE180细胞:shEn1#1组和shEn1#2组的迁移细胞数分别为(66.67±12.66)和(71.33±11.02)个, 均低于shNC组的(334.67±16.56)个(均P<0.001);KYSE450细胞:shEn1#1组和shEn1#2组的迁移细胞数分别为(112.33±14.57)和(54.33±5.51)个, 均低于shNC组的(253.33±21.03)个(均P<0.001)]。裸鼠皮下移植瘤实验Objective To explore the function and mechanism of transcription factor En1 in esophageal squamous cell carcinoma(ESCC).Methods The correlations of En1 with prognosis were analyzed using the overall survival data of 9397 pan-cancer patients and progression-free survival data of 4349 pan-cancer patients from The Cancer Genome Atlas(TCGA)database.The En1 expression data in 53 and 155 cases of ESCC and their paired adjacent tissues were from Gene Expression Omnibus(GEO)database and National Genomics Data Center-Genome Sequence Archive(NGDC-GSA)database.Lentivirus was used to generate En1 stable knockout cell lines KYSE180 and KYSE450.The proliferation ability of the cells was detected by cell counting kit 8 and clone formation assay.The migration ability of the cells was detected by Transwell assay.The effect of En1 on the proliferation of ESCC was detected by xenograft experiment in BALB/c-nu/nu mice.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expressions of En1,glioma-associated oncogene family zinc finger 1(GLI1),glioma-associated oncogene family zinc finger 2(GLI2)and smoothened(SMO).Results Pan-cancer data from TCGA showed that patients with low En1 expression had longer overall survival and progression-free survival than patients with high En1 expression(P<0.001).Data from GEO and GSA databases also showed a high expression level of En1 in ESCC tissues compared with paired tissues(P<0.001).Proliferation was inhibited after knockout of En1 in KYSE180 and KYSE450 cells(P<0.001).The colony formation numbers decreased.The colony formation numbers of KYSE180 cells in the shEn1#1 group and the shEn1#2 group were 138.33±23.07 and 127.00±19.70,respectively,significantly lower than that of the shNC group 340.67±12.06(P<0.001).The colony formation numbers of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were 65.33±2.52 and 9.00±3.00,respectively,significantly lower than that of the shNC group 139.00±13.00(P<0.001).The migration numbers was inhibited after

关 键 词：食管鳞状细胞癌 En1 HEDGEHOG通路 细胞增殖 细胞迁移 

分 类 号：R735.1[医药卫生—肿瘤]