肺腺癌中TCN1的表达及其对肺腺癌细胞增殖和迁移侵袭能力的影响  

Expression of TCN1 in lung adenocarcinoma and its effects on proliferation,migration and invasion abilities of lung adenocarcinoma cells

作　　者：黄媛媛[1] 任心怡 何杰[1] 张维[1] 黄娜[1] 姜轶[1] Huang Yuanyuan;Ren Xinyi;He Jie;Zhang Wei;Huang Na;Jiang Yi(Department of Pulmonary and Critical Care Medicine,the First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院第一附属医院呼吸与危重症医学科,成都610500

出　　处：《国际呼吸杂志》2022年第19期1464-1473,共10页International Journal of Respiration

基　　金：2018年四川省医学科研青年创新课题(Q18041);2017年度成都医学院第一附属医院专项科学研究基金(CYFY2017GLPHX02)。

摘　　要：目的探讨钴胺素转运蛋白1(TCN1)在肺腺癌中的表达及其与肺腺癌患者预后的关系,以及TCN1对肺腺癌细胞增殖和迁移侵袭能力的影响。方法采用非随机抽样的方法选取2020年7月至2021年7月在成都医学院第一附属医院行肺腺癌切除术的患者20例,收集20对经组织病理学证实的肺腺癌样本和距离肿瘤组织至少5 cm的正常肺组织样本。免疫组织化学染色分析TCN1在肺腺癌组织及癌旁正常肺组织中的表达情况,实时荧光定量聚合酶链式反应(RT-qPCR)和蛋白质印迹法(Western blot)检测肺腺癌细胞系及正常肺上皮细胞系内TCN1表达情况。下载TCGA数据库中肺腺癌数据,分析TCN1与肺腺癌患者生存预后的关系,使用加权基因共表达网络分析与TCN1关系最密切的模块,用R 3.6.1软件对该模块内的基因进行基因本体注释(GO)和京都基因和基因组百科全书(KEGG)分析,以TCN1的中位值为截断值,将TCGA数据库中的肺腺癌样本分为TCN1高表达组和低表达组,采用GSEA 4.2.3软件分析预测高表达TCN1的肿瘤组织样本内基因富集的信号通路。选用A549肺腺癌细胞系为研究对象,运用噻唑蓝(MTT)和Transwell实验检测敲减TCN1后对A549细胞的增殖和迁移侵袭的影响,采用Western blot检测敲减TCN1后A549肺腺癌细胞内上皮间充质转化(EMT)相关蛋白的表达情况。结果免疫组织化学染色显示肺腺癌患者肺腺癌组织中TCN1染色为阳性。RT-qPCR和Western blot结果显示,与正常肺上皮细胞系BEAS-2B相比,肺腺癌细胞系A549、LC-2、HCC515和PC14中TCN1普遍高表达(P值均<0.05)。对TCGA数据库中肺腺癌的生存资料进行分析显示TCN1高表达的患者总生存期更短(HR=1.6,P=0.0026)。富集分析结果显示WGCNA筛选出的与TCN1表达最相关的模块内的基因主要涉及胞外基质组成、胞外结构组成、细胞基质黏附等生物学过程以及黏着斑、细胞外基质受体相互作用等相关通路。GSEA软件�Objective To explore the expression of transcobalamin I(TCN1)in lung adenocarcinoma and its relations with prognosis of lung adenocarcinoma patients,and effects of TCNl on proliferation,migration,and invasion abilities of lung adenocarcinoma cells.Methods A non-random sampling method was conducted,2o lung adenocarcinoma samples were selected from The First Affiliated Hospital of Chengdu Medical College;meanwhile,20 normal lung tissue samples which were histopathologically confirmed and met the criteria of at least 5 cm from the tumor tissue were collected.The expressions of TCN1 in lung adenocarcinoma tissues and adjacent normal lung tissues were analyzed by immunohistochemical staining,and the expressions of TCN1 in lung adenocarcinoma cells and normal lung epithelial cell lines were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot.Lung adenocarcinoma data from the TCGA database were downloaded to analyze relations between TCNl and survival prognosis of patients with lung adenocarcinoma.The module with the closest relations between the weighted gene co-expression network analysis(WGCNA)and TCN1 was used,genes in the module were subject to gene ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis by means of R 3.6.1 software.The median value of TCN1 was used as a truncated value,lung adenocarcinoma samples from TCGA database were divided into high TCN1 expression group and low TCN1 expression group.GSEA 4.2.3 software was applied to analyze and predict the signaling pathway for gene enrichment in tumor tissue samples with high TCN1 expression.A549 lung adenocarcinoma cells were selected as the research objects.Methyl thiazolyl tetrazolium(MTT)and Transwell assay were used to detect the effects after knocking down TCNl on the proliferation,migration,and invasion of A549 cells,and Western blotting was applied to detect the expression of epithelial-to-mesenchymal transition(EMT)-related proteins in A549 lung adenocarcinoma cells after knocking down TCN

关 键 词：肺腺癌 细胞增殖 计算生物学 钴胺素转运蛋白1 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]