ASGR1在肝细胞癌中的意义及机制研究  

Study on the significance and mechanism of ASGR1 in hepatocellular carcinoma

作　　者：李倩玉 郭文韵 钱逸斐 李松玲 朱子俊 刘艳丰 LI Qianyu;GUO Wenyun;QIAN Yifei;LI Songling;ZHU Zijun;LIU Yanfeng(Renji-Med X Clinical Stem Cell Research Center,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China;School of Biomedical Engineering&Med-X Research Institute,Shanghai Jiao Tong University,Shanghai 200030,China;Department of Liver Surgery,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属仁济医院临床干细胞研究中心,上海200127 [2]上海交通大学生物医学工程学院Med-X研究院,上海200030 [3]上海交通大学医学院附属仁济医院肝外科,上海200127

出　　处：《上海交通大学学报（医学版）》2023年第9期1107-1114,共8页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(82073190);上海交通大学医学院“双百人”项目(20221704)。

摘　　要：目的·探究去唾液酸糖蛋白受体1(asialoglycoprotein receptor 1,ASGR1)在肝细胞癌(hepatocellular carcinoma,HCC)中的意义及潜在机制。方法·通过R语言分析癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中ASGR1在肝癌患者中的表达情况并绘制相关生存曲线。利用人类蛋白质图谱(The Human Protein Atlas,HPA)数据库获得人体正常肝组织和肝癌组织的免疫组织化学(immunohistochemistry,IHC)数据来分析ASGR1的蛋白表达情况。利用流体动力学尾静脉注射(hydrodynamic tail vein injection,HTVI)递送方法,在免疫完全的小鼠肝脏中敲除Asgr1探究其在体内的致瘤功能,并通过蛋白免疫印迹法(Western blotting,WB)验证基因敲除效率。利用R语言进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析及相关性分析,利用基因探针富集(Gene Set Enrichment Analysis,GSEA)软件进行GSEA hallmark相关通路分析,利用实时荧光定量PCR(quantitative real-time PCR,qPCR)在小鼠肝癌组织中验证糖酵解关键基因表达水平。结果·ASGR1在肝癌组织中显著低表达,在肝癌患者中ASGR1的低表达与患者较差的总体生存期(overall survival,OS)、无疾病间隔(disease free interval,DFI)、无进展间隔期(progression free interval,PFI)和疾病特异性生存期(disease specific survival,DSS)相关;肿瘤分级程度越高的肝癌患者ASGR1基因表达水平越低。人体正常肝组织ASGR1蛋白的表达显著高于肝癌组织。在免疫完全的肝细胞癌小鼠模型中,小鼠内源性Asgr1敲除可增加肝组织中肿瘤结节的大小和数量。TCGA数据库中ASGR1低表达组肝癌患者富集到多条癌症及代谢相关通路,ASGR1表达与部分糖酵解关键基因表达呈负相关,Asgr1敲除组的小鼠肝癌组织中糖酵解水平高于对照组,提示ASGR1低表达很可能促进肝癌的生长发展,加强代谢重编程促进肿瘤的合成代谢发展。结论·ASGR1在肝癌患者中�Objective·To explore the significance and mechanism of asialoglycoprotein receptor 1(ASGR1)in hepatocellular carcinoma.Methods·The expression of ASGR1 in patients with liver cancer in The Cancer Genome Atlas(TCGA)database was analyzed by R language and the related survival curves were drawn.The Human Protein Atlas(HPA)database was used to obtain the immunohistochemistry(IHC)data of normal human liver tissue and liver cancer tissue to analyze the protein expression of ASGR1.By using the hydrodynamic tail vein injection(HTVI)delivery method,Asgr1 was knocked out in the liver of fully immune mice to explore its tumorigenic function in vivo.Gene knockout efficiency was verified by Western blotting(WB).The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and correlation analysis were performed by using R language.The GSEA hallmark correlation pathway analysis was performed by using Gene Set Enrichment Analysis(GSEA)software.The expression level of key genes of glycolysis in mouse liver cancer tissue was verified by quantitative real-time PCR(qPCR).Results·ASGR1 was significantly low-expressed in liver cancer tissue,and the low expression of ASGR1 in liver cancer patients was associated with poorer overall survival(OS),disease-free interval(DFI),progression-free interval(PFI),and diseasespecific survival(DSS).The higher the degree of tumor grade,the lower the expression level of ASGR1 in patients with liver cancer.Immunohistochemistry showed that the protein expression of ASGR1 in normal human liver tissue was significantly higher than that in liver cancer tissue.In an immunocompetent mouse model of hepatocellular carcinoma,knockout of endogenous Asgr1 in mice increased the size and number of tumor nodules in liver tissue.In the TCGA database,patients with liver cancer in the ASGR1 lowexpression group were enriched in multiple cancer and metabolic pathways.The expression of ASGR1 was negatively correlated with some key genes of glycolysis.The level of glycolysis in liver cancer tissues of mic

关 键 词：去唾液酸糖蛋白受体1(ASGR1) 肝细胞癌 流体动力学尾静脉注射 治疗靶点 

分 类 号：R735.7[医药卫生—肿瘤]