胃癌患者血清红桂木凝集素结合蛋白的分析  

Analysis of Artocarpus lingnanensislectin binding protein in the serum of patients with gas⁃tric cancer

作　　者：谢彬彬 刘奕 李璐[1] 尹利君[1] 廖烈君 熊灏[2] 曾麒燕[1,3] Xie Binbin;Liu Yi;Li Lu;Yin Lijun;Liao Liejun;Xiong Hao;Zeng Qiyan(Department of Biochemistry and Molecular Biology;Key Laboratory of Biological Molecular Medicine Research,Guangxi Medical University,Nanning 530021,China;Department of Laboratory Medicine,Guangxi Health Vocational&Technical College,Nanning 530021,China)

机构地区：[1]广西医科大学生物化学与分子生物学教研室,南宁530021 [2]广西卫生职业技术学院医技系,南宁530021 [3]广西高校生物分子医学研究重点实验室,南宁530021

出　　处：《广西医科大学学报》2021年第5期917-922,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81160366);广西科学研究与技术开发计划项目资助(No.桂科攻10124001A-23)。

摘　　要：目的:寻找胃癌患者血清中的差异蛋白,以期为胃癌诊断提供依据。方法:应用生物素-链霉亲和素磁珠结合红桂木凝集素(ALL)后分别与正常人和胃癌患者血清共孵育,经pull-down实验、蛋白质银染、液相色谱串联质谱(LC-MS/MS)鉴定,并结合数据库分析差异蛋白的表达情况。结果:银染分析显示,胃癌组与正常组之间存在明显差异蛋白条带。从正常血清和胃癌血清中分别鉴定到1个和41个ALL结合蛋白。将ALL结合蛋白与基因表达谱数据动态分析(GEPIA)数据库中差异表达最显著的1000个胃癌相关蛋白进行交集,得到8个共有蛋白,其中免疫球蛋白重链恒定区γ2(IGHG2)、载脂蛋白E(APOE)、免疫球蛋白κ恒定区(IGKC)、免疫球蛋白κ可变区2D-28(IGKV2D-28)和Serpin家庭成员1(SERPINA1)在胃癌组织显著高表达(P<0.05),除SERPINA1外,其它4个蛋白的表达水平在胃癌不同病理分期中存在显著性差异(P<0.05)。间-α-胰蛋白酶抑制剂重链H4(ITIH4)、维生素D结合蛋白(GC)和血清类黏蛋白1(ORM1)蛋白在胃癌组织中显著低表达(P<0.05),其中ITIH4的表达水平与患者生存率相关(P<0.05)。结论:ALL结合蛋白IGHG2、APOE、IGKC、IGKV2D-28、SERPINA1、ITIH4、GC和ORM1有望成为胃癌诊断的血清标志物。Objective:To search for the differential expression proteins in serum of patients with gastric cancer,andprovide a scientific basis for the diagnosis of gastric cancer.Methods:Serum samples were incubated with Artocarpus lingnanensis lectin(ALL)binding-biotin-streptavidin magnetic beads,and followed by pull-down experiment,protein silver staining,liquid chromatography tandem mass spectrometry(LC-MS/MS)identification,and the expression of ALL-binding proteins in gastric cancer was analyzed using Gene Expression Profiling Interactive Analysis(GEPIA)database.Results:The silver staining results showed that there were significant differences in protein bands between the gastric cancer group and the normal group.One and forty-one ALL-binding proteins were identified from normal human serum and gastric cancer patient serum,respectively.Venn diagram analysis showed the overlap of proteins between ALL-binding proteins and 1,000 kinds of proteins with the most significant differential expression in gastric cancer in GEPIA database,which included IGHG2,APOE,IGKC,IGKV2D-28,SERPINA1,ITIH4,GC and ORM1.GEPIA database analysis found that the expression of IGHG2,APOE,IGKC,IGKV2D-28 and SERPINA1 were significantly higher in gastric cancer tissues than that in control group(P<0.05),and showed significantly differential expression in different pathological statuses of gastric cancer except SERPINA1(P<0.05).On the contrary,the expression of ITIH4,GC and ORM1 were significantly lower in gastric cancer tissues than that in control group(P<0.05),and the level of ITIH4 was correlated with the overall survival of gastric cancer patients(P<0.05).Conclusion:ALL-binding proteins,IGHG2,APOE,IGKC,IGKV2D-28,SERPINA1,ITIH4,GC and ORM1 are expected to be serum markers for the diagnosis of gastric cancer.

关 键 词：胃癌 红桂木凝集素 质谱 数据库 血清标志物 

分 类 号：R735.2[医药卫生—肿瘤]