摘要
目的评估自噬相关蛋白5(autophagy-related protein 5,ATG5)在结肠癌中的表达及其与临床病理特征的关系,分析ATG5对结肠癌细胞迁移及侵袭能力的影响。方法运用在线分析工具cProsite分析ATG5 mRNA以及ATG5蛋白在结肠癌组织及癌旁正常组织中的表达水平。使用Kaplan-Meier Plotter分析ATG5表达水平对结肠癌患者预后的影响。免疫组织化学法检测100例结肠癌组织中ATG5表达,χ^(2)和Fisher's检验分析ATG5表达水平与临床病理特征的相关性。转染质粒或小干扰过表达或敲低ATG5后,Western blotting检测转染效率及自噬相关标记物微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3)的变化,Transwell实验评估过表达或敲低ATG5后结肠癌细胞的迁移及侵袭能力。使用LinkedOmics数据库分析结肠癌中与ATG5相关的差异表达基因,并进行基因本体(gene ontology,GO)分析和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)分析。结果结肠癌组织中ATG5 mRNA及蛋白的表达水平均低于癌旁正常组织(P<0.001)。低表达ATG5的结肠癌患者无复发生存期明显短于高表达ATG5的结肠癌患者(P<0.001)。免疫组化结果表明,结肠癌组织中ATG5的表达水平与淋巴结转移相关,低表达ATG5的结肠癌患者更容易发生淋巴结转移(P=0.027)。Western blotting及Transwell实验显示在SW1116细胞中过表达ATG5后,LC3Ⅱ/LC3Ⅰ增加,细胞的迁移侵袭能力减弱(P_(ATG5)=0.001;P_(LC3Ⅱ/LC3Ⅰ)=0.04;P_(迁移)<0.001,P_(侵袭)<0.001);反之,在DLD1细胞中敲低ATG5后,LC3Ⅱ/LC3Ⅰ降低,细胞的迁移侵袭能力增加(P_(ATG5#1)=0.021,P_(ATG5#2)<0.001;P_(LC3Ⅱ/LC3Ⅰ#1)=0.013,P_(LC3Ⅱ/LC3Ⅰ#2)=0.02;P迁移<0.001,P侵袭<0.001)。ATG5相关的差异表达基因富集分析结果显示,结肠癌中ATG5可能通过影响DNA损伤反应、染色质重排及Notch信号通路等途径影响结肠癌的转移。结论ATG5在结肠癌中低表达,并与患者淋巴结�
Objective To evaluate the expression of autophagy-related protein 5(ATG5)in colon cancer and its relationship with clinicopathological features,and to analyze the effect of ATG5 on the migration and invasion ability of colon cancer cells.Methods The expression levels of ATG5 mRNA and ATG5 protein in colon cancer tissues and adjacent normal tissues were analyzed using the online analysis tool cProsite.Kaplan-Meier Plotter was used to analyze the effect of ATG5 expression level on the prognosis of colon cancer patients.The expression of ATG5 in 100 colon cancer tissues was detected by immunohistochemistry,and the correlation between the expression level of ATG5 and clinicopathological features was analyzed by χ^(2) and Fisher's test.ATG5 was overexpressed or knocked down in colon cancer cells by transfecting plasmid or small interfering RNA(siRNA),and then the transfection efficiency and changes of microtubule-associated protein light chain 3(LC3)were detected by Western blotting.Transwell assay was used to evaluate migration and invasion ability of colon cancer cells after overexpression or knockdown.Differentially expressed genes associated with ATG5 in colon cancer were analyzed using the Linked Omics database,and gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were performed.Results The expression levels of ATG5 mRNA and protein in colon cancer tissues were lower than those in adjacent normal tissues(P<0.001).The recurrence-free survival of colon cancer patients with low ATG5 expression was significantly shorter than those with high ATG5 expression(P<0.001).Immunohistochemistry showed that the expression level of ATG5 in colon cancer tissues was correlated with lymph node metastasis,and low expression of ATG5 in colon cancer patients was more likely to develop lymph node metastasis(P=0.027).Western blotting and Transwell experiments showed that after overexpression of ATG5 in SW1116 cells,the expression of LC3Ⅱ/LC3Ⅰincreased,and the migration and invasion ability of ce
作者
刘爱静
李雁儒
高惠茹
段伟丽
李培龙
李娟
杜鲁涛
王传新
LIU Aijing;LI Yanru;GAO Huiru;DUAN Weili;LI Peilong;LI Juan;DU Lutao;WANG Chuanxin(Department of Clinical Laboratory,The Second Hospital of Shandong University,Jinan 250033,Shandong,China;Department of Clinical Laboratory,Qilu Hospital of Shandong University,Jinan 250012,Shandong,China)
出处
《山东大学学报（医学版）》
CAS
北大核心
2024年第4期14-23,共10页
Journal of Shandong University:Health Sciences
基金
山东省重点研发计划(2021ZLGX02)。
关键词
自噬相关蛋白5
转移
自噬
结肠癌
微管相关蛋白轻链3
Autophagy-related protein 5
Tumor metastasis
Autophagy
Colon cancer
Microtubule-associated protein light chain 3
