TRIP13通过同源重组通路提高肺腺癌细胞的放射抗性  

TRIP13 Enhances Radioresistance of Lung Adenocarcinoma Cells through the Homologous Recombination Pathway

作　　者：葛舒童 谷润川 杨雄涛 许长丹 王诗杰 朱广迎 Shutong GE;Runchuan GU;Xiongtao YANG;Changdan XU;Shijie WANG;Guangying ZHU(Department of Radiation Oncology,Peking University China-Japan Friendship School of Clinical Medicine,Beijing 100029,China;China-Japan Friendship Institute of Clinical Medicine,Beijing 100029,China;Department of Oncology,Changping District Hospital,Beijing 102202,China;Department of Radiation Oncology,China-Japan Friendship Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100029,China)

机构地区：[1]北京大学中日友好临床医学院放射肿瘤科,北京100029 [2]中日友好临床研究所,北京100029 [3]北京市昌平区医院肿瘤科,北京102202 [4]中国医学科学院北京协和医学院中日友好医院放射肿瘤科,北京100029

出　　处：《中国肺癌杂志》2024年第1期1-12,共12页Chinese Journal of Lung Cancer

基　　金：中日友好医院人才引进科研启动基金项目(No.2016-RC-4)资助。

摘　　要：背景与目的放疗是非小细胞肺癌(non-small cell lung cancer,NSCLC)最常用的治疗手段之一。然而,一部分肿瘤细胞对放射线的不敏感是放疗疗效差、患者预后不良的重要原因之一,探究放射抵抗背后的深层机制是解决这一临床难题的关键。本研究旨在寻找与肺腺癌(lungadenocarcinoma,LUAD)放射抵抗相关的分子,初步经数据库筛选锁定甲状腺素受体结合因子13(thyroid hormone receptor interactor 13,TRIP13)为主要研究对象,并探索TRIP13是否与LUAD的放射抵抗有关及具体机制,以期为临床接受放疗的LUAD患者的联合治疗提供理论依据和潜在靶点。方法选取基因表达综合数据库(Gene Expression Omnibus,GEO)中的GSE18842、GSE19188和GSE33532共3个数据集,借助R 4.1.3软件分别筛选3个数据集中差异表达的基因(|logFC|>1.5,P<0.05),之后使用Venn diagram找出在3个数据集中共有的差异表达基因。随后,借助STRING在线工具和Cytoscape软件,对筛选出来的差异基因进行蛋白质相互作用分析和模块分析,借助Kaplan-Meier Plotter数据库对各基因进行生存预后分析,并确定TRIP13基因作为后续主要研究分子。随后,采用亚致死性剂量照射法对人LUAD细胞系H292进行多次X射线照射,以构建具有放射抗性的细胞系H292DR。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验和克隆形成实验验证H292DR细胞的放射抗性能力。Western blot检测H292细胞和H292DR细胞中TRIP13蛋白的表达水平。使用小干扰RNA(small interfering RNA,siRNA)沉默H292DR细胞中TRIP 13蛋白的表达并进行Western blot检测。观察TRIP13沉默后H292DR细胞的克隆形成能力和迁移能力,随后检测共济失调-毛细血管扩张突变(ataxia telangiectasia mutated,ATM)蛋白等与同源重组密切相关的蛋白的表达水平变化。结果经多个GEO数据集筛选、外部数据集的验证以及生存分析发现,TRIP13在LUAD中高表达,并与接受过放疗的LUAD患者的�Background and objective Radiation therapy is one of the most common treatments for non-small cell lung cancer(NSCLC).However,the insensitivity of some tumor cells to radiation is one of the major reasons for the poor efficacy of radiotherapy and the poor prognosis of patients,and exploring the underlying mechanisms behind radioresistance is the key to solving this clinical challenge.This study aimed to identify the molecules associated with radioresistance in lung adenocarcinoma(LUAD),identified thyroid hormone receptor interactor 13(TRIP13)as the main target initially,and explored whether TRIP13 is related to radioresistance in LUAD and the specific mechanism,with the aim of providing theoretical basis and potential targets for the combination therapy of LUAD patients receiving radiotherapy in the clinic.Methods Three datasets,GSE18842,GSE19188 and GSE33532,were selected from the Gene Expression Omnibus(GEO)database and screened for differentially expressed genes(|log FC|>1.5,P<0.05)in each of the three datasets using the R 4.1.3 software,and then Venn diagram was used to find out the differentially expressed genes common to the three datasets.The screened differential genes were then subjected to protein-protein interaction(PPI)analysis and module analysis with the help of STRING online tool and Cytoscape software,and survival prognosis analysis was performed for each gene with the help of Kaplan-Meier Plotter database,and the TRIP13 gene was identified as the main molecule for subsequent studies.Subsequently,the human LUAD cell line H292 was irradiated with multiple X-rays using a sub-lethal dose irradiation method to construct a radioresistant cell line,H292DR.The radioresistance of H292DR cells was verified using cell counting kit-8(CCK-8)assay and clone formation assay.The expression levels of TRIP13 in H292 and H292DR cells were measured by Western blot.Small interfering RNA(siRNA)was used to silence the expression of TRIP13 in H292DR cells and Western blot assay was performed.The clone formation ability

关 键 词：肺肿瘤 放射抵抗 TRIP13蛋白 同源重组 

分 类 号：R734.2[医药卫生—肿瘤]