ARID1B基因缺失促进NSCLC细胞的增殖、迁移和侵袭能力的研究  

ARID1B Gene Deletion Promotes the Proliferation,Migration and Invasion of NSCLC Cells

作　　者：朱琳琳[1,2,3] 张绪超 Linlin ZHU;Xuchao ZHANG(Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510080,China;The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510515,China;Guangdong Provincial Key Laboratory of Translational Medicine in Lung Cancer,Medical Research Center,Guangdong Provincial People's Hospital(Guangdong Academy of Medical Sciences),Southern Medical University,Guangzhou 510080,China)

机构地区：[1]南方医科大学附属广东省人民医院(广东省医学科学院),广州510080 [2]南方医科大学第二临床医学院,广州510515 [3]南方医科大学附属广东省人民医院(广东省医学科学院),医学研究部,广东省肺癌转化医学重点实验室,广州510080

出　　处：《中国肺癌杂志》2025年第3期165-175,共11页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金项目(No.82173202)资助。

摘　　要：背景与目的SWI/SNF染色质重塑复合物(switch/sucrose nonfermentable chromatin-remodeling complex)的异常与多种癌症密切相关,ARID1B(AT-rich interaction domain 1B)是SWI/SNF复合物的核心亚基之一。ARID1B基因突变或拷贝数缺失与DNA损伤反应受损、染色质可及性改变有关。然而,ARID1B缺失是否影响非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞的增殖、迁移和侵袭能力及其分子机制仍缺乏系统研究。本研究旨在揭示ARID1B基因缺失对NSCLC细胞恶性表型的调控作用及其分子机制。方法通过公共数据库分析ARID1B基因与肺癌患者预后之间的相关性以及其在肺癌组织中的表达水平。CRISPR/Cas9(clustered regularly interspaced short palindromic repeat)技术构建ARID1B基因稳定敲除(knockout,KO)的细胞株。采用平板克隆实验检测细胞增殖情况,Transwell细胞迁移、侵袭实验检测细胞迁移能力变化。RNA-Seq进行差异基因的表达、富集分析。利用蛋白印迹(Western blot,WB)验证ARID1B基因敲除效果,检测上皮间充质转化(epithelial-mesenchymal transition,EMT)标志物、促分裂素原活化蛋白激酶(mitogen-activated protein kinases,MAPK)信号通路相关蛋白变化。构建裸鼠成瘤模型,并比较对照和ARID1B缺失细胞的成瘤能力。结果低ARID1B表达与肺癌患者不良预后有显著关联,其总生存期较短。ARID1B在肺癌细胞中的表达水平较正常细胞显著降低。ARID1B缺失型细胞增殖、迁移和侵袭能力增强。动物实验中,ARID1B基因缺失组成瘤速度加快。RNA-Seq结果进行富集分析可见差异基因主要在MAPK、磷脂酰肌醇3-激酶/蛋白激酶B(phosphoinositide 3-kinase/protein kinase B,PI3K/Akt)等信号通路富集。WB证明ARID1B基因缺失细胞E-cadherin、N-cadherin、Vimentin表达发生变化,MAPK、p-MAPK表达增加。结论成功建立A549-ARID1B KO和PC9-ARID1B KO细胞株,ARID1B缺失细胞株在体外、体内生物学行为水平和转录组测序Background and objective Abnormalities of the switch/sucrose nonfermentable(SWI/SNF)chroma-tin-remodeling complex are closely related to various cancers,and ARID1B(AT-rich interaction domain 1B)is one of the core subunits of the SWI/SNF complex.Mutations or copy number deletions of the ARID1B gene are associated with impaired DNA damage response and altered chromatin accessibility.However,whether ARID1B deficiency affects the proliferation,migration and invasion abilities of non-small cell lung cancer(NSCLC)cells and its molecular mechanisms remain poorly understood.This study aims to reveal the regulatory role of ARID1B gene deletion on the malignant phenotype of NSCLC cells and its molecular mechanism.Methods Online databases were used to analyze the relationship between ARID1B and the prognosis of patients with lung cancer,and the expression levels of ARID1B in lung cancer tissues.The CRISPR/Cas9(clustered regularly interspaced short palindromic repeat)technology was employed to construct stable ARID1B gene knockout(KO)cell lines.The plate colony formation assay was used to detect cell proliferation,and the Transwell cell migration and invasion assays were used to detect changes in cell migration ability.RNA-Seq was utilized for the expression and enrichment analysis of differentially expressed genes.Western blot(WB)was used to verify the knockout effect of the ARID1B gene and to detect the expression changes of epithelial-mesenchymal transition(EMT)markers and mitogen-activated protein kinases(MAPK)signaling pathway-related proteins.Nude mouse tumor models were constructed and the tumorigenic abilities of control and ARID1B-deficient cells were compared.Results Patients with low ARID1B expression have poor overall survival.ARID1B is differentially expressed in lung cancer and normal tissues,and its expression level being lower in cancer cells.ARID1B-deficient cells had significantly enhanced in vitro prolifera-tion,migration and invasion abilities.In animal experiments,the tumor formation speed of ARID1B gene

关 键 词：ARID1B 肺肿瘤 增殖 迁移 MAPK 转录组学分析 

分 类 号：R73[医药卫生—肿瘤]